EP3979802A1 - Compositions et procédés d'amélioration de la santé des plantes et de lutte contre les maladies des plantes - Google Patents

Compositions et procédés d'amélioration de la santé des plantes et de lutte contre les maladies des plantes

Info

Publication number
EP3979802A1
EP3979802A1 EP20750436.6A EP20750436A EP3979802A1 EP 3979802 A1 EP3979802 A1 EP 3979802A1 EP 20750436 A EP20750436 A EP 20750436A EP 3979802 A1 EP3979802 A1 EP 3979802A1
Authority
EP
European Patent Office
Prior art keywords
weight ratio
active ingredient
plant
ingredient weight
acre
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP20750436.6A
Other languages
German (de)
English (en)
Inventor
David Joseph INGHAM
Vincent James Spadafora
Mathias TWIZEYIMANA
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
AgBiome Inc
Original Assignee
AgBiome Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by AgBiome Inc filed Critical AgBiome Inc
Publication of EP3979802A1 publication Critical patent/EP3979802A1/fr
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/27—Pseudomonas
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/12—Powders or granules
    • A01N25/14—Powders or granules wettable
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/34—Nitriles
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/48—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with two nitrogen atoms as the only ring hetero atoms
    • A01N43/50—1,3-Diazoles; Hydrogenated 1,3-diazoles
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/48—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with two nitrogen atoms as the only ring hetero atoms
    • A01N43/54—1,3-Diazines; Hydrogenated 1,3-diazines
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/64—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with three nitrogen atoms as the only ring hetero atoms
    • A01N43/647—Triazoles; Hydrogenated triazoles
    • A01N43/653—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles

Definitions

  • the invention relates to bacterial strains and populations for controlling plant disease and/or improving an agronomic trait of interest in a plant test
  • Plant diseases are responsible for significant agricultural losses. Effects can range from mild symptoms to catastrophic plant damage, which can lead to major economic and social consequences.
  • compositions and methods for controlling plant disease and/or for improving at least one agronomic trait of interest in a plant comprise a population of biocontrol agents or bacterial strains that control one or more pathogens that cause plant disease and/or improve at least one agronomic trait of interest.
  • the biological agents or bacterial strains can be used as an inoculant for plants.
  • Methods for growing a plant susceptible to plant disease and methods and compositions for controlling plant disease are also provided. Further provided are methods and compositions of increasing disease resistance in plants. Methods and compositions for improving plant health and/or improving at least one agronomic trait of interest are also provided.
  • compositions and methods for improving at least one agronomic trait of interest and/or improving plant health and/or for controlling one or more plant diseases are provided.
  • the compositions and methods described herein comprise a combination (applied simultaneously or sequentially) of a biocontrol agent and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • a biological agent, biocontrol agent, bacterial strain, modified bacterial strain, modified biological agent, or modified biocontrol agent or active variant thereof are used herein to describe a microorganism that is used to control disease-causing plant pathogens and/or improve at least one agronomic trait of interest and/or improve plant health.
  • Combination (applied simultaneously or sequentially) of a biocontrol agent with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole can provide an improvement in at least one agronomic trait of interest and/or improving plant health and/or for controlling one or more plant diseases are provided.
  • lower amounts of the biocontrol agent and/or the synthetic fungicide can provide an improvement in at least one agronomic trait of interest and/or plant health and/or can control one or more plant diseases than when applying the standard amounts of the biocontrol agent and synthetic fungicide independently.
  • the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole with the biocontrol agents listed herein produce at least an additive effect, and in particular embodiments a synergistic effect, that allows for use of a lower amount of the biocontrol agent and/or synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) than the suggested or commonly used amount for application.
  • synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difeno
  • lower amounts of biocides can be added to crops or plants in order to increase disease control, reduce chemical residues, reduce pathogen resistance, and increase product usage base acres in multiple crops for the bacterial strain and synthetic fungicide.
  • lower rates of the biocontrol agent and the synthetic fungicide can provide a synergistic effect (i.e., greater than additive or superadditive).
  • a“synthetic fungicide” is a chemical substance that can exert a controlling effect on one or more fungi or fungal-like organismby chemical means, leading to the death of the fungus or fungal- like organism or its spores or inhibition of the growth of the fungus or the fungal-like organism.
  • the synthetic fungicide of the presently disclosed compositions and methods is tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • Tetraconazole, tebuconazole, flutriafol, and difenoconazole are triazoles.
  • Azoxystrobin is a methoxyacrylate strobilurin antifungal agent.
  • Chlorothalonil is an aromatic fungicide.
  • biocontrol agents or bacterial strains are provided which can be used in combination with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to control one or more plant disease and/or improve at least one agronomic trait of interest and/or improve plant health.
  • Such bacterial strains include AIP1620 and AIP050999.
  • AIP1620 is a Pseudomonas strain that has been selected for glyphosate tolerance.
  • AIP050999 is a Pseudomonas strain that has been selected for glufosinate tolerance.
  • Biocontrol agents further comprise CGA267356 (ATTC Accession no. 55169) described in U.S. Patent No. 5,348,742.
  • Cell populations comprising one or more of AIP1620, AIP050999, and CGA267356 are provided.
  • various bacterial strains and/or the pesticidal compositions provided herein comprise as an active ingredient (a) a cell population comprising one or more of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof along with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • AIP1620 was deposited with the Patent Depository of the National Center for Agricultural
  • isolated encompasses a bacterium, spore, or other entity or substance, that has been (1) separated from at least some of the components with which it was associated when initially produced (whether in nature or in an experimental setting), and/or (2) produced, prepared, purified, and/or manufactured by the hand of man.
  • Isolated bacteria may be separated from at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or more of the other components with which they were initially associated.
  • a substance is “pure” if it is substantially free of other components.
  • the terms “purify,” “purifying” and “purified” refer to a bacterium, spore, or other material that has been separated from at least some of the components with which it was associated either when initially produced or generated (e.g., whether in nature or in an experimental setting), or during any time after its initial production.
  • a bacterium or spore or a bacterial population or a spore population may be considered purified if it is isolated at or after production, such as from a material or environment containing the bacterium or bacterial population or spore, and a purified bacterium or bacterial population or spore may contain other materials up to about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or above about 90% and still be considered purified.
  • purified bacteria or spores and bacterial populations or spore populations are more than about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% pure.
  • a culture of bacteria contains no other bacterial species in quantities to be detected by normal bacteriological techniques.
  • population is intended a group or collection that comprises two or more (i.e., 10, 100, 1,000, 10,000, lxlO 6 , lxlO 7 , or lxlO 8 or greater).
  • compositions are provided herein that comprise a population of at least one bacterial strain.
  • the population of at least one of a bacterial strain comprises a concentration of at least about 10 3 CFU/ml to about 10 5 CFU/ml, 10 3 CFU/ml to about 10 4 CFU/ml, 10 3 CFU/ml to about 10 6 CFU/ml, 10 4 CFU/ml to about 10 8 CFU/ml, 10 5 CFU/ml to about 10 11 CFU/ml, about 10 5 CFU/ml to about 10 10 CFU/ml, about 10 5 CFU/ml to about 10 12 CFU/ml, about 10 5 CFU/ml to about 10 6 CFU/ml, about 10 6 CFU/ml to about 10 7 CFU/ml, about 10 7 CFU/ml to about 10 8 CFU/ml, about 10 8 CFU/ml to about 10 9 CFU/ml, about 10 9 CFU/ml, about 10 9 CFU/ml, about 10 9 CFU/ml, about 10 9 CFU/ml, about 10 9 CFU
  • the concentration of the bacterial strain provided herein or active variant thereof comprises at least about 10 2 CFU/ml, at least about 10 3 CFU/ml, at least about 10 4 CFU/ml, at least about 10 5 CFU/ml, at least about 10 6 CFU/ml, at least about 10 7 CFU/ml, at least about 10 8 CFU/ml, at least about 10 9 CFU/ml, at least about 10 10 CFU/ml, at least about 10 11 CFU/ml, or at least about 10 12 CFU/ml or equivalent measure of bacterial concentration.
  • the population of at least one bacterial strain comprises a concentration of at least about 10 3 CFU/g to about 10 4 , 10 3 CFU/g to about 10 5 CFU/g, CFU/g, 10 3 CFU/g to about 10 6 CFU/g, 10 4 CFU/g to about 10 8 CFU/g, 10 5 CFU/g to about 10 11 CFU/g, about 10 5 CFU/g to about 10 10 CFU/g, about 10 5 CFU/g to about 10 12 CFU/g, about 10 5 CFU/g to about 10 6 CFU/g, about 10 6 CFU/g to about 10 7 CFU/g, about 10 7 CFU/g to about 10 8 CFU/g, about 10 8 CFU/g to about 10 9 CFU/g, about 10 9 CFU/g to about 10 10 CFU/g, about 10 10 CFU/g to about 10 11 CFU
  • the concentration of the bacterial strain provided herein or active variant thereof comprises at least about 10 2 CFU/g, at least about 10 3 CFU/g, at least about 10 4 CFU/g, at least about 10 5 CFU/g, at least about 10 6 CFU/g, at least about 10 7 CFU/g, at least about 10 8 CFU/g, at least about 10 9 CFU/g, at least about 10 10 CFU/g, at least about 10 11 CFU/g, or at least about 10 12 CFU/g or equivalent measure of bacterial concentration.
  • the bacterial concentration of a given solid or liquid composition or formulation can be expressed in CFU/g or CFU/mU, respectively, or by an equivalent measure of bacterial concentration using any methods described herein.
  • a measure of bacterial concentration that is equivalent to CFU can be expressed in terms of cells/g or cells/mU if using epifluore scent measurements or pg of pyrrolnitrin per g of bacteria when using pyrrolnitrin measurements as a reporter metabolite.
  • the concentration of the bacterial strain can be lower in a composition intended for application as a diluted mixture when compared to the concentration of the bacterial strain in a composition intended for application as a diluted mixture in the absence of the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole).
  • compositions comprising a bacterial strain i.e., at least one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof, and/or at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole can further comprise an agriculturally acceptable carrier.
  • Agriculturally acceptable carrier is intended to include any material that facilitates application of a composition to the intended subject (i.e, a plant or plant part susceptible to a plant disease of interest (i.e., Asian Soybean Rust (ASR), or any other disease disclosed herein or a plant or plant part for improving an agronomic trait of interest)).
  • Carriers used in compositions for application to plants and plant parts are preferably non phytotoxic or only mildly phytotoxic.
  • a suitable carrier may be a solid, liquid or gas depending on the desired formulation.
  • carriers include polar or non-polar liquid carriers such as water, mineral oils and vegetable oils. Additional carriers are disclosed elsewhere herein.
  • compositions and/or forumulations comprising the bacterial strain, or variant thereof, disclosed herein can be combined with at least one synthetic fungicide composition as disclosed herein, as a pre-mix or, if appropriate, not combined until immediately prior to use (i.e, tank mix).
  • a pre-mix formulation can comprise 1 to 95 percent, of the desired ingredients (bacterial composition or synthetic fungicide), 99 to 5 percent, of a solid or liquid adjuvant (including, for example, a solvent such as water), where the auxiliaries 0.5 to 40 percent, based on the pre-mix formulation.
  • compositions and/or forumulations comprising the bacterial strain, or variant thereof, disclosed herein can be packaged with a at least one synthetic fungicide composition as disclosed herein.
  • a single container can comprise a packaged bacterial composition or formulation and a separately packaged synthetic fungicide formulation.
  • the compositions and/or forumulations comprising the bacterial strain, or variant thereof, and the synthetic chemistry disclosed herein can be provided in a single container as a pre-mix formulation or as a blended formulation.
  • compositions comprising at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and/or active variants of AIP1620, AIP050999, and/or CGA267356.
  • Variants of AIP1620, AIP050999, and/or CGA267356 will retain the ability to control one or more plant diseases (i.e., reduce disease severity and/or reduce disease development) and/or control one or more plant pathogens when applied in combination with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • plant diseases i.e., reduce disease severity and/or reduce disease development
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • variants of AIP1620, AIP050999, and/or CGA267356 will retain the ability to control one or more fungal plant diseases and/or plant disease caused by a fungal-like pathogen and/or one or more fungal pathogens and/or one or more fungal-like pathogen in combination with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • variants will retain the ability to control ASR when applied in combination (simultaneously or sequentially) with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • variants will retain the ability to synergistically control one or more fungal plant diseases, plant diseases caused by fungal-like pathogens, and/or one or more fungal pathogens or fungal-like pathogens in combination with at least one synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutrafiol, tebuconazole, chlorothalonil, or difenoconazole).
  • synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutrafiol, tebuconazole, chlorothalonil, or difenoconazole.
  • Active variants of the various bacterial strains provided herein include, for example, any isolate or mutant of AIP1620, AIP050999, and CGA267356.
  • the bacterial strain is applied in combination (simultaneously or sequentially) with at least one synthetic fungicide (which is a synthetic biocide) selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • a biocide is a chemical substance that can exert a controlling effect on an organism by chemical or biological means.
  • Biocides include pesticides, such as fungicides; herbicides; insecticides, other crop protection chemicals, and the like.
  • a bacterial strain (e.g., AIP1620, AIP050999, and CGA267356, or an active variant thereof) is compatible with a biocide when the bacterial strain is able to survive, germinate into a vegetative cell, and/or reproduce in the presence of an effective amount of a biocide of interest.
  • methods can be undertaken to modify the bacterial strain to impart the compatibility of interest. Such methods to produce modified bacterial strains include both selection techniques and/or transformation techniques.
  • Methods and compositions disclosed herein can include a bacterial strain and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole,
  • chlorothalonil and difenoconazole, along with another biocide to further increase plant health and/or control plant disease.
  • modified bacterial strain is intended a population wherein the strain has been modified (by selection and/or transformation) to have one or more additional traits of interest.
  • the modified bacterial strain comprises any one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof.
  • the modified bacterial strain is compatible with a biocide of interest, including but not limited to, resistance to a herbicide, fungicide, pesticide, or other crop protection chemical.
  • the modified biocide-resistant strains have the same identification characteristics as the original sensitive strain except they are significantly more resistant to the particular herbicide, fungicide, pesticide, or other crop protection chemical. Their identification is readily possible by comparison with characteristics of the known sensitive strain. Thus, isolated populations of modified bacterial strains are provided.
  • An increase in resistance to a biocide refers to the ability of an organism (i.e., bacterial cell or spore) to survive and reproduce following exposure to a dose of the biocide (e.g, herbicide, fungicide, pesticide, or other crop protection chemical) that would normally be lethal to the unmodified organism or would substantially reduce growth of the unmodified organism.
  • the increase in resistance to a biocide is demonstrated in the presence of an agriculturally effective amount of the biocide, which is the amount needed to exert its intended effect (e.g., controlling weed(s), controlling pest(s), plant pathogen(s)).
  • compositions provided herein include selected or engineered bacterial strains and modified populations of bacterial strains. These bacterial strains or modified bacterial strains can be used as an inoculant for plants. They can also be applied as a spray application directly to the aerial parts of plants, and can be mixed with the herbicide or other chemical to which they have been modified to become tolerant.
  • active variants of the bacterial strains disclosed herein include for example, a modified strain, such that the active variants act in combination with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, fhitriafol, tebuconazole, chlorothalonil, and difenoconazole to control a plant disease and further are able to grow in the presence of at least one biocide.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, fhitriafol, tebuconazole, chlorothalonil, and difenoconazole to control a plant disease and further are able to grow in the presence of at least one biocide.
  • Recombinant bacterial strains having resistance to an herbicide, fungicide, pesticide, or other crop protection chemical can be made through genetic engineering techniques and such engineered or recombinant bacterial strains grown to produce a modified population of bacterial strains.
  • a recombinant bacterial strain is produced by introducing polynucleotides into the bacterial host cell by transformation. Methods for transforming microorganisms are known and available in the art. See, generally, Hanahan, D (1983) Studies on transformation of Escherichia coli with plasmids J. Mol. Biol. 166, 557-77; Seidman,
  • Transformation may occur by natural uptake of naked DNA by competent cells from their environment in the laboratory.
  • cells can be made competent by exposure to divalent cations under cold conditions, by electroporation, by exposure to polyethylene glycol, by treatment with fibrous nanoparticies, or other methods well known in the art.
  • Herbicide resistance genes for use in transforming a recombinant bacterial strain include, but are not limited to, fumonisin detoxification genes (U.S. Patent No. 5,792,931); acetolactate synthase (ALS) mutants that lead to herbicide resistance, in particular the sulfonylurea-type herbicides, such as the S4 and/or Hra mutations; inhibitors of glutamine synthase such as phosphinothricin or basta (e.g., bar gene); and glyphosate resistance (EPSPS gene); gluphosinate, and HPPD resistance (WO 96/38576, U.S. Patent Nos.
  • Patent No. 7,935,869; and U.S. Patent No. 8,124,846 are herein incorporated by reference.
  • the bar gene encodes resistance to the herbicide basta
  • the nptll gene encodes resistance to the antibiotics kanamycin and geneticin
  • the ALS-gene mutants encode resistance to the sulfonylurea herbicides including
  • the bacterial strains are grown in the presence of the herbicide, fungicide, pesticide, or other crop protection chemical as the selection pressure. Susceptible agents are killed while resistant agents survive to reproduce without competition. As the bacterial strains are grown in the presence of the herbicide, fungicide, pesticide, or other crop protection chemical, resistant bacterial strains successfully reproduce and become dominant in the population, becoming a modified population of bacterial strains. Methods for selecting resistant strains are known and include U.S. Patent Nos. 4,306,027 and 4,094,097, herein incorporated by reference.
  • the active variant of the bacterial strain comprising a modified population of bacterial strains will have the same identification characteristics as the original sensitive strain except they are significantly more tolerant to the particular herbicide, fungicide, pesticide, or other crop protection chemical. Thus, their identification is readily possible by comparison with characteristics of the known sensitive strain.
  • the active variants of the bacterial strain(s) AIP1620, AIP050999, and CGA267356 include strains that are closely related to any of the disclosed strains by employing the Bishop MLST method of organism classification as defined in Bishop et al (2009) BMC Biology 7(1)1741-7007-7-3.
  • an active variant of a bacterial strain disclosed herein includes a bacterial strain that falls within at least a 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%. 94%, 95%, 96%, 97%, 98%, 98.5%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% sequence cut off employing the Bishop method of organism classification as set forth in Bishop et al. (2009) BMC Biology 7(1)1741-7007-7-3, which is herein incorporated by reference in its entirety.
  • Active variants of the bacteria identified by such methods will retain the ability to improve at least one agronomic trait when applied with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to a plant, plant part, or an area of cultivation, including for example, reducing plant disease severity and/or reducing plant disease development.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to a plant, plant part, or an area of cultivation, including for example, reducing plant disease severity and/or reducing plant disease development.
  • the active variant of the bacterial strain(s) disclosed herein include strains that are closely related to any of the disclosed strains on the basis of the Average Nucleotide Identity (ANI) method of organism classification.
  • ANI Average Nucleotide Identity
  • an active variant of bacterial strain AIP1620, A1P050999, and CGA267356 disclosed herein includes a bacterial stain that falls within at least a 90%, 95%, 96%, 97%, 97.5%, 98%, 98.5%, 98.8%, 99%, 99.5%, or 99.8% sequence cut off employing the ANI method of organism classification as set forth in Konstantinidis, K.T., et al, (2005) PNAS USA 102(7):2567-72, which is herein incorporated by reference in its entirety.
  • Active variants of the bacteria identified by such methods will retain the ability to improve at least one agronomic trait when applied (simultaneously or sequentially) with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to a plant, plant part, or an area of cultivation, including for example, reducing plant disease severity and/or reducing plant disease development.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to a plant, plant part, or an area of cultivation, including for example, reducing plant disease severity and/or reducing plant disease development.
  • the active variants of the isolated bacterial strain(s) disclosed herein include strain(s) that are closely related to any of the above strains (for example, closely related to AIP1620, AIP050999, and CGA267356) on the basis of 16S rDNA sequence identity. See Stackebrandt E, et al, “Report of the ad hoc committee for the re-evaluation of the species definition in bacteriology,” IntJSyst Evol Microbiol. 52(3): 1043-7 (2002) regarding use of 16S rDNA sequence identity for determining relatedness in bacteria.
  • the at least one strain is at least 95% identical to any of the above strains on the basis of 16S rDNA sequence identity, at least 96% identical to any of the above strains on the basis of 16S rDNA sequence identity, at least 97% identical to any of the above strains on the basis of 16S rDNA sequence identity, at least 98% to any of the above strains on the basis of 16S rDNA sequence identity, at least 98.5% identical to any of the above strains on the basis of 16S rDNA sequence identity, at least 99% identical to any of the above strains on the basis of 16S rDNA sequence identity, at least 99.5% to any of the above strains on the basis of 16S rDNA sequence identity or at least 100% to any of the above strains on the basis of 16S rDNA sequence identity.
  • Active variants of the bacteria identified by such methods will retain the ability to improve at least one agronomic trait when applied (simultaneously or sequentially) with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole in an effective amount to a plant, plant part, or an area of cultivation, including for example, reducing plant disease severity and/or reducing plant disease development.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole in an effective amount to a plant, plant part, or an area of cultivation, including for example, reducing plant disease severity and/or reducing plant disease development.
  • MinHash distance metric is a comparison method that defines thresholds for hierarchical classification of microorganisms at high resolution and requires few parameters and steps (Ondov et al. (2016) Genome Biology 17: 132). Mash distance strongly corresponds to Average Nucleotide Identity method (ANI) for hierarchical classification (See, Konstantinidis, K.T. et al. (2005) PNAS USA
  • an ANI of 97% is approximately equal to a Mash distance of 0.03, such that values put forth as useful classification thresholds in the ANI literature can be directly applied with the Mash distance.
  • Active variants of the bacterial strain(s) disclosed herein include strains that are closely related to any of AIP1620, AIP050999, and CGA267356 on the basis of the Minhash (Mash) distance between complete genome DNA sequences.
  • an active variant of a bacterial strain disclosed herein includes bacterial strains having a genome within a Mash distance of less than about 0.015 to the disclosed strains.
  • an active variant of a bacterial strain disclosed herein includes a distance metric of less than about 0.005, 0.010, 0.015, 0.020, 0.025, or 0.030.
  • a genome, as it relates to the Mash distance includes both bacterial chromosomal DNA and bacterial plasmid DNA.
  • the active variant of a bacterial strain has a genome that is above a Mash distance threshold to the disclosed strains that is greater than dissimilarity caused by technical variance. In further instances, the active variant of a bacterial strain has a genome that is above a Mash distance threshold to the disclosed strains that is greater than dissimilarity caused by technical variance and has a Mash distance of less than about 0.015. In other instances, the active variant of a bacterial strain has a genome that is above a Mash distance threshold to the disclosed strains that is greater than dissimilarity caused by technical variance and has a Mash distance of less than about 0.005, 0.010, 0.015, 0.020, 0.025, or 0.030.
  • “above technical variation” means above the Mash distance between two strains caused by errors in the genome assemblies provided the genomes being compared were each DNA sequenced with at least 20X coverage with the Illumina HiSeq 2500 DNA sequencing technology and the genomes are at least 99% complete with evidence for contamination of less than 2%. While 20X coverage is an art recognized term, for clarity, an example of 20X coverage is as follows: for a genome size of 5 megabases (MB), 100 MB of DNA sequencing from the given genome is required to have 20X sequencing coverage on average at each position along the genome. There are many suitable collections of marker genes to use for genome completeness calculations including the sets found in Campbell et al. (2013) PNAS USA 110(14):5540-45, Dupont et al.
  • Genomes of sufficient quality for comparison must be produced.
  • a genome of sufficient quality is defined as a genome assembly created with enough DNA sequence to amount to at least 20X genome coverage using Illumina HiSeq 2500 technology. The genome must be at least 99% complete with contamination of less than 2% to be compared to the claimed microbe’s genome.
  • Genomes are to be compared using the Minhash workflow as demonstrated in Ondov et al. (2016) Genome Biology 17: 132, herein incorporated by reference in its entirety. Unless otherwise stated, parameters employed are as follows:“sketch” size of 1000, and“k-mer length” of 21.
  • the bacterial strains provided herein can be formulated as a cell paste, wettable powders, a cell pellet, dusts, granules, a slurry, a dry powder, a spray dried formulation, an agglomerated formulation, a fluidized bed agglomerated formulation, aqueous or oil based liquid products, and the like.
  • Such formulations will comprise the bacteria provided herein or an active variant thereof in addition to carriers and other agents.
  • the formulation will further comprise at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and any carrier or other agent necessary to formulate the synthetic fungicide with the bacterial strains provided herein (i.e., AIP1620, AIP050999, and CGA267356).
  • the bacterial / synthetic chemistry formulation can comprise a co-pack, a blended formulation or a pre-mix formulation. The formulations can be used in a variety of methods as disclosed elsewhere herein.
  • the bacterial strains disclosed herein and the active variants thereof can be formulated to include at least one or more of an extender, a solvent, spontaneity promoters, carriers, emulsifiers, dispersants, frost protectants, thickeners, and/or adjuvants.
  • the synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole
  • the synthetic fungicide can be formulated to include at least one or more of an extender, a solvent, spontaneity promoters, carriers, emulsifiers, dispersants, frost protectants, thickeners, and/or adjuvants.
  • Examples of typical formulations include water-soluble liquids (SL), emulsifiable concentrates (EC), emulsions in water (EW), flowable liquid formulations (F; a formulation wherein the active ingredient is a solid that does not dissolve in either water or oil), cell pastes, dry flowable formulations, suspension concentrates (SC, SE, FS, OD), water-dispersible granules (WDG), granules (GR), spray dried formulation, wettable powders, agglomerated formulation, fluidized bed agglomeration formulation, and capsule concentrates (CS); WDG; GR; BB; SG; ZC these and other possible types of formulation are described, for example, by Crop Life International and in Pesticide Specifications, Manual on development and use of FAO and WHO specifications for pesticides, FAO Plant Production and Protection Papers - 173, prepared by the FAO/WHO Joint Meeting on Pesticide Specifications, 2004, ISBN: 9251048576.
  • the formulations may comprise active agrochemical compounds other than
  • the formulations or application forms of the various bacterial strains or active variants thereof and/or synthetic fungicidial chemistries can comprise, but are not limited to, auxiliaries, such as extenders, solvents, spontaneity promoters, carriers, emulsifiers, dispersants, frost protectants, biocides, solid carriers, surfactants, thickeners and/or other auxiliaries, such as adjuvants.
  • auxiliaries such as extenders, solvents, spontaneity promoters, carriers, emulsifiers, dispersants, frost protectants, biocides, solid carriers, surfactants, thickeners and/or other auxiliaries, such as adjuvants.
  • An adjuvant in this context is a component which enhances the biological effect of the formulation, without the component itself having a biological effect.
  • adjuvants are agents which promote the retention, spreading, attachment to the leaf surface, or penetration.
  • Non-limiting extenders are, for example, water, polar and nonpolar organic chemical liquids, for example from the classes of the aromatic and non-aromatic hydrocarbons (such as paraffins, alkyl benzenes, alkylnaphthalenes, chlorobenzenes), the alcohols and polyols (which, if appropriate, may also be substituted, etherified and/or esterified), the ketones (such as acetone, cyclohexanone), esters (including fats and oils) and (poly)ethers, the unsubstituted and substituted amines, amides, lactams (such as N-alkylpyrrolidones) and lactones, the sulphones and sulphoxides (such as dimethyl sulphoxide).
  • aromatic and non-aromatic hydrocarbons such as paraffins, alkyl benzenes, alkylnaphthalenes, chlorobenzenes
  • non-limiting liquid solvents are: aromatics such as xylene, toluene or alkylnaphthalenes, chlorinated aromatics and chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffins, for example petroleum fractions, mineral and vegetable oils, alcohols such as butanol or glycol and also their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents such as dimethylformamide and dimethyl sulphoxide, and also water.
  • aromatics such as xylene, toluene or alkylnaphthalenes
  • chlorinated aromatics and chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride
  • Non-limiting solvents are, for example, aromatic hydrocarbons, such as xylene, toluene or alkylnaphthalenes, for example, chlorinated aromatic or aliphatic hydrocarbons, such as chlorobenzene, chloroethylene or methylene chloride, for example, aliphatic hydrocarbons, such as cyclohexane, for example, paraffins, petroleum fractions, mineral and vegetable oils, alcohols, such as methanol, ethanol, isopropanol, butanol or glycol, for example, and also their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, for example, strongly polar solvents, such as dimethyl sulphoxide, and water.
  • aromatic hydrocarbons such as xylene, toluene or alkylnaphthalenes
  • chlorinated aromatic or aliphatic hydrocarbons such as chlor
  • Non-limiting examples of suitable carriers include, for example, ammonium salts and ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic minerals, such as finely divided silica, alumina and natural or synthetic silicates, resins, waxes and/or solid fertilizers. Mixtures of such carriers may likewise be used.
  • ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth
  • ground synthetic minerals such as finely divided silica, alumina and natural or synthetic silicates, resins, waxes and/or solid fertilizers. Mixtures of such carriers may likewise be used.
  • Carriers suitable for granules include the following: for example, crushed and fractionated natural minerals such as calcite, marble, pumice, sepiolite, dolomite, and also synthetic granules of inorganic and organic meals, and also granules of organic material such as sawdust, paper, coconut shells, maize cobs, and tobacco stalks.
  • Liquefied gaseous extenders or solvents may also be used.
  • extenders or carriers which at standard temperature and under standard pressure are gaseous, examples being aerosol propellants, such as halogenated hydrocarbons, and also butane, propane, nitrogen and carbon dioxide.
  • aerosol propellants such as halogenated hydrocarbons, and also butane, propane, nitrogen and carbon dioxide.
  • emulsifiers and/or foam-formers, dispersants or wetting agents having ionic or nonionic properties, or mixtures of these surface-active substances are salts of polyacrylic acid, salts of lignosulphonic acid, salts of phenolsulphonic acid or naphthalenesulphonic acid, polycondensates of ethylene oxide with fatty alcohols or with fatty acids or with fatty amines, with substituted phenols
  • sulphosuccinic esters preferably alkylphenols or arylphenols
  • taurine derivatives preferably alkylta urates
  • phosphoric esters of polyethoxylated alcohols or phenols preferably alkylta urates
  • phosphoric esters of polyethoxylated alcohols or phenols preferably alkylta urates
  • phosphoric esters of polyethoxylated alcohols or phenols preferably alkylta urates
  • phosphoric esters of polyethoxylated alcohols or phenols preferably alkylta urates
  • phosphoric esters of polyethoxylated alcohols or phenols preferably alkylta urates
  • phosphoric esters of polyethoxylated alcohols or phenols preferably alkylta urates
  • phosphoric esters of polyethoxylated alcohols or phenols preferably alkylta urates
  • auxiliaries that may be present in the formulations and in the application forms derived from them include colorants such as inorganic pigments, examples being iron oxide, titanium oxide, Prussian Blue, and organic dyes, such as alizarin dyes, azo dyes and metal phthalocyanine dyes, and nutrients and trace nutrients, such as salts of iron, manganese, boron, copper, cobalt, molybdenum, and zinc.
  • colorants such as inorganic pigments, examples being iron oxide, titanium oxide, Prussian Blue, and organic dyes, such as alizarin dyes, azo dyes and metal phthalocyanine dyes, and nutrients and trace nutrients, such as salts of iron, manganese, boron, copper, cobalt, molybdenum, and zinc.
  • Stabilizers such as low-temperature stabilizers, preservatives, antioxidants, light stabilizers or other agents which improve chemical and/or physical stability may also be present. Additionally present may be foam-formers or defoamers.
  • formulations and application forms derived from them may also comprise, as additional auxiliaries, stickers such as carboxymethylcellulose, natural and synthetic polymers in powder, granule or latex form, such as gum arabic, polyvinyl alcohol, polyvinyl acetate, and also natural phospholipids, such as cephalins and lecithins, and synthetic phospholipids.
  • additional auxiliaries include mineral and vegetable oils.
  • auxiliaries present in the formulations and the application forms derived from them.
  • additives include fragrances, protective colloids, binders, adhesives, thickeners, thixotropic substances, penetrants, retention promoters, stabilizers, sequestrants, complexing agents, humectants and spreaders.
  • the active compounds may be combined with any solid or liquid additive commonly used for formulation purposes.
  • Suitable retention promoters include all those substances which reduce the dynamic surface tension, such as dioctyl sulphosuccinate, or increase the viscoelasticity, such as hydroxypropylguar polymers, for example.
  • Suitable penetrants in the present context include all those substances which are typically used in order to enhance the penetration of active agrochemical compounds into plants.
  • Penetrants in this context are defined in that, from the (generally aqueous) application liquor and/or from the spray coating, they are able to penetrate the cuticle of the plant and thereby increase the mobility of the active compounds in the cuticle. This property can be determined using the method described in the literature (Baur et ah, 1997, Pesticide Science 51: 131-152).
  • Examples include alcohol alkoxylates such as coconut fatty ethoxylate (10) or isotridecyl ethoxylate (12), fatty acid esters such as rapeseed or soybean oil methyl esters, fatty amine alkoxylates such as tallowamine ethoxylate (15), or ammonium and/or phosphonium salts such as ammonium sulphate or diammonium hydrogen phosphate, for example.
  • alcohol alkoxylates such as coconut fatty ethoxylate (10) or isotridecyl ethoxylate (12)
  • fatty acid esters such as rapeseed or soybean oil methyl esters
  • fatty amine alkoxylates such as tallowamine ethoxylate (15)
  • ammonium and/or phosphonium salts such as ammonium sulphate or diammonium hydrogen phosphate, for example.
  • compositions and formulations disclosed herein can comprise an amount of a bacterial strain, such as AIP1620, AIP050999, and CGA267356, or active variant of any thereof, or a combination of cells from any one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof.
  • the bacterial strain can occur in any formulation type of interest, including, for example in a wettable powder, spray dried formulation, dry flowable formulation, agglomerated formulation, fluidized bed agglomerated forumation, or in a cell paste.
  • the bacterial strain and the synthetic chemistry are formulated separately and packaged as a co-pack or a blend, while in other embodiments, the bacterial strain and the synthetic chemistry are formulated as a pre-mix formulation.
  • compositions and formulations disclosed herein comprise an amount of a bacterial strain, such as AIP1620, AIP050999, and CGA267356, or active variant of any thereof, and at least one synthetic fungicide at various active ingredient weight ratios.
  • active ingredient weight ratio refers to the quantitative relation between the weights of each of two active ingredients within a
  • compositions are those that have the ability to improve at least one agronomic trait when applied in an effective amount to a plant, plant part, or an area of cultivation, including for example, reducing plant disease severity and/or reducing plant disease
  • the active ingredients of the presently disclosed compositions are at least one synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) and/or a bacterial strain, such as AIPI620, AIP050999, and CGA267356, or active variant of any thereof.
  • synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole
  • a bacterial strain such as AIPI620, AIP050999, and CGA267356, or active variant of any thereof.
  • the total weight of the bacterial strain can be used to calculate the active ingredient weight ratio.
  • the active ingredient weight ratio can be measured using any method known in the art, including weighing the amount of a dry formulation comprising one or both of the active ingredients to obtain the formulation weight and then calculating the weight of each active ingredient based on the reported, known, or calculated percentage of the active ingredient to the total formulation weight (w/w). For example, 2 pounds of a dry formulation that is 50% w/w of the active ingredient chlorothalonil comprises 1 pound of chlorothalonil.
  • the weight of each active ingredient can be calculated by measuring the total volume of the formulation and then calculating the weight of each active ingredient based on the reported, known, or calculated percentage of the active ingredient to the total formulation volume (w/v).
  • Percent weight per volume is defined as the grams of solute in 100 ml of a solution.
  • 100 ml of a liquid composition that is 50% w/v of the active ingredient azoxystrobin comprises 50 g of azoxystrobin.
  • One non-limiting method that can be used to measure the weight of a bacterial strain (cells, spores, etc.) or an active variant thereof in liquid culture includes pelleting the cells using, for example, a centrifuge, in order to remove any liquid and then weighing the pelleted cells. The effectiveness of a given bacterial cell weight can be expressed by measuring the culturability, viability or activity (e.g., measuring a reporter metabolite such as pyrrolnitrin), as discussed elsewhere herein.
  • the culturablity of a bacterial strain such as AIP1620, AIP050999, and CGA267356, or active variant of any thereof, in a composition or formulation can be quantitated by measuring the number of colony forming units per gram or per ml of the formulation.
  • the composition or formulation comprises a concentration of the bacterial strain of at least about 10 4 to about 10 12 CFU/gram,
  • 10 4 to about 10 10 CFU/gram at least about 10 5 CFU/gram to about 10 11 CFU/gram, about 10 5 CFU/gram to about 10 10 CFU/gram, about 10 5 CFU/gram to about 10 12 CFU/gram, about 10 5 CFU/gram to about 10 6 CFU/gram, about 10 6 CFU/gram to about 10 7 CFU/gram, about 10 7 CFU/gram to about 10 8 CFU/gram, about 10 8 CFU/gram to about 10 9 CFU/gram, about 10 9 CFU/gram to about 10 10 CFU/gram, about 10 10 CFU/gram to about 10 11 CFU/gram, or about 10 11 CFU/gram to about 10 12 CFU/gram.
  • 10 6 CFU/gram about 10 6 CFU/gram to about 10 7 CFU/gram
  • about 10 7 CFU/gram to about 10 8 CFU/gram about 10 8 CFU/gram to about 10 9 CFU/gram
  • about 10 9 CFU/gram to about 10 10 CFU/gram about 10 10 CFU/gram to about
  • the concentration of the bacterial strain comprises at least about 10 4 CFU/gram, at least about
  • the composition or formulation comprises a concentration of the bacterial strain of at least about 10 4 to about 10 12 CFU/mU, 10 4 to about 10 10 CFU/mU at least about 10 5 CFU/mU to about 10 11 CFU/mU, about 10 5 CFU/mU to about 10 10 CFU/mU, about 10 5 CFU/mU to about 10 12 CFU/mU, about 10 5 CFU/mU to about 10 6 CFU/mU, about 10 6 CFU/mU to about 10 7 CFU/mU, about 10 7 CFU/mU to about 10 8 CFU/mU, about 10 8 CFU/mU to about 10 9 CFU/mU, about 10 9 CFU/mU to about 10 10 CFU/mU, about 10 10 CFU/mU to about 10 11 CFU/mU, or about 10 11 CFU/mU to about 10 12 CFU/mU.
  • the concentration of the bacterial strain comprises at least about 10 4 CFU/mU, at least about 10 5 CFU/mU, at least about 10 6 CFU/mU, at least about 10 7 CFU/mU, at least about 10 8 CFU/mU, at least about 10 9 CFU/mU, at least about 10 10 CFU/mU, at least about 10 11 CFU/mU, or at least about 10 12 CFU/mU, or equivalent measure of bacterial concentration.
  • the viability of a bacterial strain such as AIP162Q, AIP050999, and CGA267356, or active variant of any thereof, in a composition or formulation can be quantitated using an epifluorescence assay in which fluorescent dyes that are specific for cells with intact membranes or disrupted membranes are utilized, such as those assays that use a SYTO 9 nucleic acid stain that fluoresces green indicating a cell has an intact membrane and propidium iodide that fluoresces red indicating a cell with a disrupted membrane that is not viable (see, for example, UIVE/DEAD ® Bac LightTM Bacterial Viability and Counting Kit from Molecular Probes; and Ivanova et al. (2010) Biotechnology &
  • the composition or formulation comprises a concentration (e.g., as measured by viability) of the bacterial strain of at least about 10 1 cells/gram to about 10 6 cells/gram, 10 2 cells/gram to about 10 5 cells/gram, 10 2 cells/gram to about 10 4 cells/gram, 10 3 cells/gram to about 10 6 cells/gram, 10 4 cells/gram to about 10 8 cells/gram, at least about 10 5 cells/gram to about 10 11 cells/gram, about 10 7 cells/gram to about 10 10 cells/gram, about 10 7 cells/gram to about 10 11 cells/gram, about 10 6 cells/gram to about 10 10 cells/gram, about 10 6 cells/gram to about 10 11 cells/gram, about 10 11 cells/gram to about 10 12 cells/gram, about 10 5 cells/gram to about 10 10 cells/gram, about 10 5 cells/gram to about 10 12 cells/gram, about 10 5 cells/gram to about 10 6 cells/gram, about 10 6 cells/gram to about 10 7 cells/gram, about 10 7 cells/gram to about 10 8 cells/gram,
  • the concentration of the bacterial strain comprises at least about 10 2 cells/gram, at least about 10 3 cells/gram, at least about 10 4 cells/gram, at least about 10 5 cells/gram, at least about 10 6 cells/gram, at least about 10 7 cells/gram, at least about 10 8 cells/gram, at least about 10 9 cells/gram, at least about 10 10 cells/gram, at least about 10 11 cells/gram, at least about 10 12 cells/gram, or at least about 10 13 cells/gram of viable cells as measured with an epifluorescence assay.
  • the amount of bacterial strain, or active variant thereof, disclosed herein can comprise a concentration of at least about 10 1 cells/mL to about 10 6 cells/mL, 10 2 cells/mL to about 10 5 cells/mL, 10 2 cells/mL to about 10 4 cells/mL, 10 3 cells/mL to about 10 6 cells/mL, 10 4 cells/mL to about 10 8 cells/mL, at least about 10 3 to about 10 9 cells/mL, at least about 10 3 to about 10 6 cells/mL, at least about 10 4 to about 10 11 cells/mL, at least about 10 5 cells/mL to about 10 11 cells/mL, about
  • 10 6 cells/mL about 10 6 cells/mL to about 10 7 cells/mL, about 10 7 cells/mL to about 10 8 cells/mL, about 10 8 cells/mL to about 10 9 cells/mL, about 10 9 cells/mL to about 10 10 cells/mL, about 10 10 cells/mL to about 10 11 cells/mL, or about 10 11 cells/mL to about 10 12 cells/mL or at least about 10 3 cells/mL, at least about 10 4 cells/mL, at least about 10 5 cells/mL, at least about 10 6 cells/mL, at least about 10 7 cells/mL, at least about 10 8 cells/mL, at least about 10 9 cells/mL, at least about 10 10 cells/mL, at least about 10 11 cells/mL, at least about 10 12 cells/mL of viable cells as measured with an epifluorescence assay.
  • the concentration of pyrrolnitrin within a composition or formulation comprising a bacterial strain can be measured as a surrogate of the viability and/or antifungal activity of the bacterial strain in the composition or formulation.
  • Pyrrolnitrin can be measured as a reporter metabolite for antifungal activity as it is co-regulated with other antifungal metabolites that are active in AIP1620, AIP050999, and CGA267356, or active variants of any thereof.
  • the presence of pyrrolnitrin is a measure of intact cells and cell concentration within a composition or formulation.
  • Pyrrolnitrin and other antifungal metabolites are retained within cells and not secreted, so measurement first requires cell lysis. Pyrrolnitrin can then be measured using any analytical chemistry method known in the art, including but not limited to, high performance liquid chromatography with ultraviolet detection (HPLC-UV) of a composition or formulation, such as that described in Hill et al. (1994) Appl Env Micro 60(1) 78-85, which is herein incorporated by reference in its entirety.
  • HPLC-UV high performance liquid chromatography with ultraviolet detection
  • compositions or formulations comprise between about 100 pg/g to 2000 pg/g, 200 pg/g to 1800 pg/g, 300 pg/g to 1500 pg/g, 300 pg/g to 1300 pg/g, 400 pg/g to 1500 pg/g, 400 pg/g to 1300 pg/g, 300 pg/g to 1000 pg/g, 400 pg/g to 1000 pg/g, 500 pg/g to 1000 pg/g, 500 pg/g to 1300 pg/g, 600 pg/g to 1000 pg/g, 600 pg/g to 1300 pg/g, 600 pg/g to 1500 pg/g, or about 300 pg/g, about 400 pg/g, about 500 pg/g, about 600 pg/g, about 700 pg/g, about 800 pg/g, about 900
  • the bacterial strain and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole in the formulations can be mixed or applied (simultaneously or sequentially) in an active ingredient weight ratio that results in at least an additive effect when applied to a plant, plant plart, or area of plant cultivation.
  • the bacterial strain, or active variant thereof can be combined with tetraconazole or flutriafol in an active ingredient weight ratio (lb/lb) of about 1 : 10 (bacterial strain: synthetic fungicide) to about 1000: 1, including but not limited to about 1: 10, about 1 :9, about 1 :8, about 1 :7, about 1 :6, about 1 :5, about 1 :4, about 1:3, about 1 :2, about 1 : 1, about 2: 1, about 3: 1, about 4: 1, about 5: 1, about 6: 1, about 7: 1, about 8: 1, about 9: 1, about 10: 1, about 15: 1, about 20: 1, about 25: 1, about 30: 1, about 40: 1, about 50: 1, about 60: 1, about 70: 1, about 80: 1, about 90: 1, about 100: 1, about 150: 1, about 200: 1, about 300: 1, about 400: 1, about 500: 1, about 600: 1, about 700: 1, about 800: 1, about 900: 1 or about 1000: 1.
  • the bacterial strain, or active variant thereof can be combined (in a formulation or applied in combination, simultaneously or sequentially) with tetraconazole or flutriafol in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain synthetic fungicide) to about 100: 1, including but not limited to about 1: 1, about 2: 1, about 5: 1, about 10: 1, about 15: 1, about 20: 1, about 25: 1, about 30: 1, about 35: 1, about 40: 1, about 45: 1, about 50: 1, about 55: 1, about 60: 1, about 61 : 1, about 62: 1, about 62.5: 1, about 63: 1, about 64: 1, about 65: 1, about 66: 1, about 67: 1, about 68: 1, about 69: 1, about 70: 1, about 75: 1, about 80: 1, about 85: 1, about 90: 1, about 95: 1, and about 100: 1.
  • the bacterial strain, or active variant thereof can be combined (in a formulation or applied in combination, simultaneously or sequentially) with azoxystrobin, tebuconazole, or difenoconazole in an active ingredient weight ratio (lb/lb) of about 1 : 10 (bacterial strain synthetic fungicide) to about 500: 1, about 1 :5 to about 100: 1, about 1 :5 to about 75: 1, about 1 :5 to about 50: 1, about 1:5 to about 20: 1, about 1 : 1 to about 100: 1, about 1 : 1 to about 75: 1, about 1: 1 to about 50: 1, about 1 : 1 to about 20: 1, including but not limited to about 1: 10, about 1 :9, about 1 :8, about 1 :7, about 1 :6, about 1 :5, about 1 :4, about 1:3, about 1:2, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9: 1, about 10: 1, about 15:
  • the bacterial strain, or active variant thereof can be combined (in a formulation or applied in combination, simultaneously or sequentially) with azoxystrobin in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain synthetic fungicide) to about 25: 1 or about 1 : 1 to about 20: 1 combined (in a formulation or applied in combination, simultaneously or sequentially) with tebuconazole in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain synthetic fungicide) to about 50:1, including but not limited to about 1:1, about 3:1, about 5:1, about 7:1, about 9:1, about 11:1, about 13:1, about 15:1, about 17:1, about 17.5:1, about 18:1, about 18.5:1, about 19:1, about 19.5:1, about 20:1, about 16:1, about 17:1, about 18:1, about 19:1, about 19.1:1, about 19.2:1, about 19.3:1, about 19.4:1, about 19.5:1, about 19.6:1, about 19.7:
  • the bacterial strain, or active variant thereof can be combined (in a formulation or applied in combination, simultaneously or sequentially) with difenconazole in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain synthetic fungicide) to about 50:1, about 1 : 1 to about 40:1, about 10: 1 to about 40: 1, or about 10: 1 to about 30:1, including but not limited to, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26:1, about 27:1, about 28:1, about 29:1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35.1:1, about 36:1, about 37:1, about 38
  • the bacterial strain, or active variant thereof can be combined (in a formulation or applied in combination, simultaneously or sequentially) with chlorothalonil in an active ingredient weight ratio (lb/lb) of about 1 : 100 and about 500: 1, including but not limited to about 1 : 100, about 1:50, about 1:10, about 1:1, about 10:1, about 50:1, about 100:1, about 150:1, about 200:1, about 250: 1, about 300: 1, about 350: 1, about 400: 1, about 450: 1, and about 500: 1.
  • active ingredient weight ratio lb/lb
  • the bacterial strain, or active variant thereof can be combined (in a formulation or applied in combination, simultaneously or sequentially) with chlorothalonil in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial strain synthetic fungicide) to about 10:1, including but not limited to, about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1.9, about 1:1.8, about 1:1.75, about 1:1.7, about 1:1.65, about 1:1.6, about 1:1.55, about 1:1.5, about 1:1.45, about 1:4, about 1:1.35, about 1:1.3, about 1:1.275, about 1:1.25, about 1:1.24, about 1:1.23, about 1:1.22, about 1:1.21, about 1:1.2, about 1:1.19, about 1:1.18, about 1:1.17, about 1:1.16, about 1:1.15, about 1:1.15, about 1:1.1, about 1:1, about 1.1:1, about 1.2:1, about 1.3:1, about 1:10 (bacterial strain synthetic
  • the bacterial strain, or active variant thereof can be combined with (in a formulation or applied in combination, simultaneously or sequentially) trifluimizole in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial strain driflumizole) to about 100: 1, including but not limited to about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 2: 1, about 3:1, about 4: 1, about 5:1, about 6: 1, about 7:1, about 8:1, about 9: 1, about 10:1, about 15:1, about 20: 1, about 25:1, about 30: 1, about 35:1, about 40: 1, about 45:1, about 50: 1, about 55:1, about 60: 1, about 65:1, about 70:1, about 75:1, about 80:1, about 85:1, about 90:1, about 95:1, or about 100:1.
  • active ingredient weight ratio lb/lb
  • the bacterial strain, or active variant thereof can be combined with (in a formulation or applied in combination, simultaneously or sequentially) with triflumizole in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial straimtriflumizole) to about 25: 1 or about 1 : 10 to about 20:1, including but not limited to about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 1.1:1, about 1.2:1, about 1.3:1, about 1.4:1, about 1.5:1, about 1.6:1, about 1.61:1, about 1.62:1, about 1.63:1, about 1.64:1, about 1.65:1, about 1.66:1, about 1.67:1, about 1.68:1, about 1.69:1, about 1.7:1, about 1.8:1, about 1.9:1, about 2:1, about 2:1, about 2.5:1, about 3:1, about 3.5:1, about 4:1, about 4.5:1, about 5:1, about 5.5:1, about 6
  • the formulations are in the form of a pre-mixture of the bacterial strain(s) and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole, each supplied as a WDG.
  • the bacterial strain and synthetic chemistry i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole,
  • chlorothalonil, or difenoconazole) in the form of WDG can be mixed in any weight ratio that results in at least an additive effect when applied to a plant, plant plart, or area of plant cultivation.
  • the bacterial strain, or active variant thereof can be combined with tetraconazole or flutriafol in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial strain synthetic fungicide) to about 1000: 1, including but not limited to about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8: 1, about 9: 1, about 10: 1, about 15:1, about 20: 1, about 25:1, about 30: 1, about 40: 1, about 50: 1, about 60: 1, about 70: 1, about 80: 1, about 90: 1, about 100: 1, about 150:1, about 200: 1, about 300: 1, about 400: 1, about 500: 1, about 600: 1, about 700: 1, about 800: 1, about 900: 1 or about 1000: 1 as WDG in a single composition.
  • active ingredient weight ratio lb/lb
  • the bacterial strain, or active variant thereof can be combined with tetraconazole or flutriafol in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain synthetic fungicide) to about 100:1, including but not limited to about 1:1, about 2:1, about 5:1, about 10:1, about 15:1, about 20:1, about 25:1, about 30:1, about 35:1, about 40:1, about 45:1, about 50:1, about 55:1, about 60: 1, about 61:1, about 62: 1, about 62.5: 1, about 63:1, about 64: 1, about 65:1, about 66: 1, about 67: 1, about 68:1, about 69:1, about 70:1, about 75:1, about 80:1, about 85:1, about 90:1, about 95:1, and about 100:1 as WDG in a single composition.
  • active ingredient weight ratio lb/lb
  • the bacterial strain, or active variant thereof can be combined with azoxystrobin, tebuconazole, or difenoconazole in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial strain: synthetic fungicide) to about 500:1, about 1:5 to about 100:1, about 1:5 to about 75:1, about 1:5 to about 50:1, about 1:5 to about 20:1, about 1:1 to about 100:1, about 1:1 to about 75:1, about 1:1 to about 50:1, or about 1:1 to about 20:1, including but not limited to about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 15:1, about 20:1, about 25:1, about 30:1, about 40: 1, about 50: 1, about 60: 1, about 70: 1, about 80: 1, about 90
  • the bacterial strain, or active variant thereof can be combined with azoxystrobin in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain synthetic fungicide) to about 25:1 or about 1:10 to about 20:1, including but not limited to about 1:1, about 1.5:1, about2:l, about 2.5:1, about 3:1, about 3.5:1, about 4:1, about 4.5:1, about 5:1, about 5.5:1, about 6:1, about 6.1:1, about 6.2:1, about 6.3:1, about 6.4:1, about 6.5:1, about 6.6:1, about 6.7:1, about 6.8:1, about 6.9:1, about 7:1, about 7.1:1, about 7.2:1, about 7.3:1, about 7.4:1, about 7.5:1, about 7.6:1, about 7.7:1, about 7.8:1, about 7.9:1, about 8:1, about 8.5:1, about 9:1, about 9.5:1, about 10:1, about 10.25:1, about 10.4:1, about 10.5:1, about 10.75:1, about 11:
  • the bacterial strain, or active variant thereof can be combined with tebuconazole in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain synthetic fungicide) to about 50:1, including but not limited to about 1:1, about 3:1, about 5:1, about 7:1, about 9:1, about 11:1, about 13:1, about 15:1, about 17:1, about 17.5:1, about 18:1, about 18.5:1, about 19:1, about 19.5:1, about 20:1, about 16:1, about 17:1, about 18:1, about 19:1, about 19.1:1, about 19.2:1, about 19.3:1, about 19.4:1, about 19.5:1, about 19.6:1, about 19.7:1, about 19.8:1, about 19.9:1, about 20:1, about 20.25:1, about 20.5:1, about 20.75:1, about 21:1, about 21.25:1, about 21.5:1, about 21.75:1, about 22:1, about 22.2:1, about 22.22:1, about 22.25: 1, about 22.5:1, about 22
  • the bacterial strain, or active variant thereof can be combined with difenconazole in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain: synthetic fungicide) to about 50:1, about 1 : 1 to about 40: 1, about 10: 1 to about 40: 1, or about 10: 1 to about 30:1, including but not limited to, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26:1, about 27:1, about 28:1, about 29:1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35.1:1, about 36:1, about 37:1, about 38:1, about 39:1, about 40:1, about 41:1,
  • the bacterial strain, or active variant thereof can be combined with chlorothalonil in an active ingredient weight ratio (lb/lb) of about 1 : 100 to about 500: 1, including but not limited to, about 1:100, about 1:50, about 1:10, about 1:1, about 10:1, about 50:1, about 100:1, about 150:1, about 200: 1, about 250: 1, about 300: 1, about 350: 1, about 400: 1, about 450: 1, and about 500: 1 as WDG in a single composition.
  • active ingredient weight ratio lb/lb
  • the bacterial strain, or active variant thereof can be combined with chlorothalonil in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial strain: synthetic fungicide) to about 10:1, including but not limited to, about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1.9, about 1:1.8, about 1:1.75, about 1:1.7, about 1:1.65, about 1:1.6, about 1:1.55, about 1:1.5, about 1:1.45, about 1:4, about 1:1.35, about 1:1.3, about 1:1.275, about 1:1.25, about 1:1.24, about 1:1.23, about 1:1.22, about 1:1.21, about 1:1.2, about 1:1.19, about 1:1.18, about 1:1.17, about 1:1.16, about 1:1.15, about 1:1.15, about 1:1.1, about 1:1, about 1.1:1, about 1.2:1, about 1.3:1, about 1.4:1, about 1.5:1, about 1.6:1, about
  • the bacterial strain, or active variant thereof can be combined with trifluimizole in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial straimtriflumizole) to about 100:1, including but not limited to about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 15:1, about 20:1, about 25:1, about 30:1, about 35:1, about 40:1, about 45:1, about 50:1, about 55:1, about 60:1, about 65:1, about 70:1, about 75:1, about 80:1, about 85:1, about 90: 1, about 95:1, or about 100: 1 as WDG in a single composition.
  • active ingredient weight ratio lb/lb
  • the bacterial strain, or active variant thereof can be combined with triflumizole in an active ingredient weight ratio (lb/lb) of about 1 : 10 (bacterial straimtriflumizole) to about 25: 1 or about 1 : 10 to about 20: 1, including but not limited to about 1: 10, about 1 :9, about 1 :8, about 1 :7, about 1:6, about 1 :5, about 1 :4, about 1 :3, about 1 :2, about 1 : 1, about 1.1 : 1, about 1.2: 1, about 1.3: 1, about 1.4: 1, about 1.5: 1, about 1.6: 1, about 1.66: 1, about 1.7: 1, about 1.8: 1, about 1.9: 1, about 2: l, about 2: l, about 2.5: 1, about 3: 1, about 3.5: 1, about 4: 1, about 4.5: 1, about 5: 1, about 5.5: 1, about 6: 1, about 6.5: 1, about 7: 1, about 7.5: 1, about 8: 1, about 8.5: 1, about 9: 1, about 9.5: 1, about
  • Water-soluble liquids, emulsifiable concentrates, emulsions in water, flowable liquid formulations, dry flowable formulations, suspension concentrates (SC, SE, FS, OD), water-dispersible granules, granules, wettable powders, spray dried formulation, agglomerated formulation, fluidized bed agglomeration formulation, cell pastes, and capsule concentrates can comprise a bacterial strain, such as AIP1620, AIP050999, and CGA267356, or active variant of any thereof.
  • the amount of the bacterial strain can comprise a concentration of the bacterial strain of at least about 10 4 to about 10 13 CFU/gram, at least about 10 5 CFU/gram to about 10 11 CFU/gram, about 10 7 CFU/gram to about 10 10 CFU/gram, about 10 7 CFU/gram to about 10 11 CFU/gram, about 10 6 CFU/gram to about 10 10 CFU/gram, about 10 6 CFU/gram to about 10 11 CFU/gram, about 10 11 CFU/gram to about 10 12 CFU/gram, about 10 5 CFU/gram to about 10 10 CFU/gram, about 10 5 CFU/gram to about 10 12 CFU/gram, about 10 5 CFU/gram to about 10 6 CFU/gram, about 10 6 CFU/gram to about 10 7 CFU/gram, about 10 7 CFU/gram to about 10 8 CFU/gram, about 10 8 CFU/gram to about 10 9 CFU/gram, about 10 9 CFU/gram to about 10 10 CFU/gram, about 10 10 CFU/gram to about 10 11 C
  • the concentration of the bacterial strain comprises at least about 10 5 CFU/gram, at least about 10 6 CFU/gram, at least about 10 7 CFU/gram, at least about 10 8 CFU/gram, at least about 10 9 CFU/gram, at least about 10 10 CFU/gram, at least about 10 11 CFU/gram, at least about 10 12 CFU/gram, or at least about 10 13 CFU/gram.
  • a “cell paste” comprises a population of cells that has been centrifuged and/or fdtered or otherwise concentrated.
  • a coated seed which comprises a seed and a coating on the seed.
  • the coating comprises a combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain, such as AIP1620, AIP050999, and CGA267356, or active variant of any thereof.
  • the coating can comprise the combination of synthetic fungicide and bacterial strain in any weight ratio disclosed herein, wherein the combination comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells.
  • the seed coating can be applied to any seed of interest (i.e., for a monocot or a dicot). Various plants of interest are disclosed elsewhere herein.
  • a seed coating can further comprise at least at least one nutrient, at least one herbicide or at least one pesticide, or at least one biocide. See, for example, US App Pub. 20040336049, 20140173979, and 20150033811.
  • the seed coating can comprise a bacterial strain, such as AIP1620, AIP050999, and CGA267356, or active variant of any thereof, along with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • At least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole can be provided in the cultivation area of the planted seed having a seed coat comprising a bacterial strain described herein.
  • compositions and formulations disclosed herein can comprise an amount of a bacterial strain, such as AIP1620, AIP050999, and CGA267356, or active variant of any thereof, along with an amount of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • the bacterial strain can be mixed with the synthetic fungicide in any amount that results in an improvement in plant health or control of a plant disease or plant pathogen, and/or improvement of an agronomic trait of interest in a plant.
  • the bacterial strain is mixed with the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) in an amount that results in at least an additive improvement in plant health or control of a plant disease or plant pathogen, and/or improvement of an agronomic trait of interest in a plant.
  • the synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole
  • the bacterial strain is mixed with the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) in an amount that results in a synergistic improvement in plant health or control of a plant disease or plant pathogen, and/or improvement of an agronomic trait of interest in a plant.
  • the synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole
  • compositions and formulations disclosed herein include a bacterial strain and/or at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole that comprises less than the suggested amount of the bacterial strain and/or the synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole when applied alone.
  • synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole when applied alone.
  • the term“suggested amount,”“standard amount,”“suggested rate,” or “standard rate” in reference to a synthetic fungicide or bacteria refers to an amount or rate that is the amount or rate that effectively controls a plant pathogen, treats or prevents a plant disease, or improves an agronomic trait of interest in a plant when used alone (i.e., not in conjunction with an additional fungicide).
  • the suggested or standard amount or rate can be the amount or rate approved for use by an applicable government agency or the amount or rate suggested by the manufacturer or listed on the label of a commercial product comprising the synthetic fungicide or bacteria.
  • the suggested amount can differ based on the particular plant pathogen being targeted or plant disease being treated or prevented or the particular agronomic trait of interest that is desired to be improved or the particular plant that the bacteria and synthetic fungicide is being applied thereto, the particular type of application (e.g., foliar, field inoculation), or the like.
  • compositions and formulations comprising at least one synthetic fungicide can comprise a bacterial strain for application at about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or about 20-70%, about 30-60%, about 30-70%, about 40-80%, about 40- 70%, about 40-60%, about 40-50%, about 50-90%, about 50-80%, about 50-70%, about 50-60%, about 60- 90%, about 60-80%, about 60-70%, about 70-90%, about 70-90%, or about 80-90% of the suggested amount for application when used alone (i.e., not in conjunction with an additional fungicide).
  • the suggested amount for application of the bacterial strain can be about 1 lb/acre, about 1.25 lb/acre, about 1.5 lb/acre, about 1.75 lb/acre, about 2 lb/acre, about 2.25 lb/acre, about 2.5 lb/acre, about 2.75 lb/acre, about 3 lb/acre, about 3.5 lb/acre, about 4 lb/acre, about 5 lb/acre, about 6 lb/acre, about 7 lb/acre, about 8 lb/acre, about 1- 10 lb/acre, or about 1-8 lb/acre.
  • a formulated product comprising AIP1620 may have 50% AIP1620 by weight or be provided in any weight ratio relative to the selected synthetic fungicide disclosed elsewhere herein.
  • the application rate of the synthetic fungicide disclosed herein can be calculated based on the rate of the bacterial strain, or active varian thereof, according to the selected active ingredient weight ratio as disclosed elsewhere herein for each selected synthetic fungicide.
  • Application of 5 lbs/acre of the 50% formulated product comprises 2.5 lbs/acre of the bacterial strain AIP1620.
  • compositions and formulation comprising a bacterial strain can comprise at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole at about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or about 20-70%, about 30-60%, about 30-70%, about 40- 80%, about 40-70%, about 40-60%, about 40-50%, about 50-90%, about 50-80%, about 50-70%, about 50- 60%, about 60-90%, about 60-80%, about 60-70%, about 70-90%, about 70-90%, or about 80-90% of the suggested amount for application when used alone.
  • synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutria
  • the standard use of bacterial strain AIP1620 is 5 lb/acre of a 50% formulation, wherein the combination comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells.
  • the bacterial strain, or active fragment thereof, and the synthetic chemistry can be formulated together in the same composition or formulated separately for mixing in a tank for application to a plant, plant part, or cultivation area of a plant.
  • formulations for mixing in a tank for application are formulated for dilution 1 : 1 with water.
  • the various formulations disclosed herein can be stable for at least 30, 40, 50, 60, 70, 80, 90, 100, 125, 150, 200, 225, 250, 275, 300, 325, 350 days, 1.5 years, 2 years or longer.
  • stable is intended that the formulation retains viable bacteria and/or retains an effective amount of a biologically active bacteria.
  • the stable formulation retains at least about 1%, about 10%, about 20%, about 30% about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% of the viability or culturability in the formulation at a given storage time point when compared to the viability or culturability produced after immediate preparation of the formulation.
  • the stable formulation retains at least about 30% to 80%, about 50% to about 80%, about 60% to about 70%, about 70% to about 80%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70% of biological activity (e.g., antifungal activity as calculated using a reporter metabolite such as pyrrolnitrin) in the formulation at a given storage time point when compared to the biological activity found in the formulation immediately after production.
  • biological activity e.g., antifungal activity as calculated using a reporter metabolite such as pyrrolnitrin
  • the stable formulation at a given storage time point retains at least about 30%, 45%, 50%, 60%, 70%, 80%, 90% of biological activity when compared to the biological activity found in the formulation immediately after production.
  • the stable formation retains any combination of the viability and biological activity noted above.
  • the formulations preferably comprise between 0.00000001 % and 98% by weight of each active compound or, with particular preference, between 0.01 % and 95% by weight of each active compound, more preferably between 0.5% and 90% by weight of each active compound, based on the weight of the formulation.
  • the active compounds content of the application forms prepared from the formulations may vary within wide ranges.
  • the active compounds concentration of the application forms may be situated typically between 0.00000001 % and 95% by weight of each active compound, preferably between 0.00001 % and 1 % by weight, based on the weight of the application form.
  • Application takes place in a customary manner adapted to the application forms.
  • the bacterial strain provided herein or an active variant thereof can be mixed or applied with a biocide in addition to at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole, such as a fungicide, insecticide, or herbicide to enhance its activity or the activity of the chemical to which it has been added.
  • a biocide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole, such as a fungicide, insecticide, or herbicide to enhance its activity or the activity of the chemical to which it has been added.
  • the combination (simultaneous or sequential application to a plant or an area of cultivation) of the bacterial strain and synthetic fungicide may exhibit an additive effect on controlling a plant pathogen or treating or preventing a plant disease or improving an agronomic trait of interest.
  • the combination of the bacterial strain and synthetic fungicide may show synergistic activity where the mixture of the two exceeds that expected from their simple additive effect.
  • the simultaneous or sequential application of the bacterial strain and synthetic fungicide to a plant or an area of cultivation results in the controlling of a plant pathogen or treatment or prevention of a plant disease or an improvement of an agronomic trait of interest wherein no such effect results when either the bacterial strain or synthetic fungicide are used alone.
  • the bacterial strain or active variant thereof is compatible with agricultural chemicals used to improve performance of biocides.
  • agricultural chemicals include safeners, surfactants, stickers, spreaders, UV protectants, and suspension and dispersal aids.
  • Safeners are chemicals that improve or modify the performance of herbicides.
  • Surfactants, spreaders, and stickers are chemicals included in agricultural spray preparations that change the mechanical properties of the spray (for example, by altering surface tension or improving leaf cuticle penetration).
  • UV protectants improve the performance of agricultural biocides by reducing degradation by ultraviolet light.
  • Suspension and dispersal aids improve the performance of biocides by altering their behavior in a spray tank.
  • methods can be undertaken to modify the bacterial strain to impart the compatibility of interest. Such methods to produce modified bacterial strains include both selection techniques and/or transformation techniques.
  • the combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and a bacterial strain, or active variant thereof, provided herein can be used to improve at least one agronomic trait of interest (e.g., reduce disease such as ASR or another fungal or fungal-like pathogen of interest).
  • compositions and formulations including at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and a bacterial strain, or active variant thereof, provided herein can be used with other pesticides for an effective integrated pest management program.
  • the biocontrol populations can be mixed with known pesticides in a manner described in WO 94/10845, herein incorporated by reference.
  • Non-limiting examples of compounds and compositions that can be added to the formulation include but are not limited to, Acetyl tributyl citrate [Citric acid, 2-(acetyloxy)-, tributyl ester]; Agar;
  • Calcium oxide silicate (Ca 3 0(Si0z t )); Calcium silicate [Silicic acid, calcium salt]; Calcium stearate
  • Glyceryl stearate [Octadecanoic acid, ester with 1,2,3-propanetriol]; Granite; Graphite; Guar gum; Gum Arabic; Gum tragacanth; Gypsum; Hematite (Fe203); Humic acid; Hydrogenated cottonseed oil;
  • Hydrogenated rapeseed oil Hydrogenated soybean oil; Hydroxyethyl cellulose [Cellulose, 2-hydroxyethyl ether]; Hydroxypropyl cellulose [Cellulose, 2-hydroxypropyl ether]; Hydroxypropyl methyl cellulose
  • Soapbark [Quillaja saponin]; Soapstone; Sodium acetate [Acetic acid, sodium salt]; Sodium alginate;
  • Vermiculite Vermiculite; Vinegar (maximum 8% acetic acid in solution); Vitamin C [L-Ascorbic acid]; Vitamin; Walnut flour; Walnut shells; Wheat; Wheat flour; Wheat germ oil; Whey; White mineral oil (petroleum);
  • kits can comprise instructions for use.
  • the kit comprises a bacterial strain, such as AIP1620, AIP050999, and
  • the premixed composition is a water dispersible granule.
  • the kit provides at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole in a spatially separated arrangement from the bacterial strain, such as AIP1620, AIP050999, and CGA267356, or an active variant of any thereof.
  • the bacterial strain or active variants thereof are provided as a wettable powder.
  • the synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole is provided as a water dispersible granule.
  • tebuconazole is provided as a dry flowable (DF).
  • azoxystrobin, flutriafol, chlorothalonil, triflumazole, and difenconazole is provided as a flowable liquid formulation (F).
  • the bacterial strain and/or the synthetic fungicide are comprised within a vessel(s), such as a box, bag, or bottle.
  • the kit comprises the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) and the bacterial strain, such as AIP1620, AIP050999, and CGA267356, or an active variant of any thereof in a single vessel (e.g., box, bag, or bottle) with a partition between two compartments of the vessel, wherein the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) is in one compartment and the bacterial strain is in the other compartment.
  • the two compartments can each have a lid that can be opened or closed independently of the other.
  • the partition between the two compartments is removable to allow mixing of the two
  • the bacterial strains or modified bacterial strains or active variants thereof provided herein can be applied (simultaneously or sequentially) with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to any plant species in order to improve an agronomic trait of interest.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to any plant species in order to improve an agronomic trait of interest.
  • the bacterial strains disclosed herein and/or the synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole
  • the synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole
  • the synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole
  • Agronomic traits of interest include any trait that improves plant health or commercial value.
  • Non-limiting examples of agronomic traits of interest include increase in biomass, increase in drought tolerance, thermal tolerance, herbicide tolerance, drought resistance, insect resistance, f mgus resistance, virus resistance, bacteria resistance, male sterility, cold tolerance, salt tolerance, increased yield, enhanced nutrient use efficiency, increased nitrogen use efficiency, increased tolerance to nitrogen stress, increased fermentable carbohydrate content, reduced lignin content, increased antioxidant content, enhanced water use efficiency, increased vigor, increased germination efficiency, earlier or increased flowering, increased biomass, altered root-to-shoot biomass ratio, enhanced soil water retention, or a combination thereof.
  • the agronomic trait of interest includes an altered oil content, altered protein content, altered seed carbohydrate composition, altered seed oil composition, and altered seed protein composition, chemical tolerance, cold tolerance, delayed senescence, disease resistance, drought tolerance, ear weight, growth improvement, health enhancement, heat tolerance, herbicide tolerance, herbivore resistance, improved nitrogen fixation, improved nitrogen utilization, improved root architecture, improved water use efficiency, increased biomass, increased root length, increased seed weight, increased shoot length, increased yield, increased yield under water-limited conditions, kernel mass, kernel moisture content, metal tolerance, number of ears, number of kernels per ear, number of pods, nutrition enhancement, pathogen resistance, pest resistance, photosynthetic capability improvement, salinity tolerance, stay-green, vigor improvement, increased dry weight of mature seeds, increased fresh weight of mature seeds, increased number of mature seeds per plant, increased chlorophyll content, increased number of pods per plant, increased length of pods per plant, reduced number of wilted leaves per plant, reduced number of severely wilted leaves per plant, increased number of
  • the bacterial strain or active variant thereof provided herein can be applied (simultaneously or sequentially) with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to decrease or reduce the level of a plant pest.
  • fungicides include but are not limited to, insects, fungi, bacteria, nematodes, acarids, protozoan pathogens, animal-parasitic liver flukes, and the like.
  • the bacterial strain or active variant thereof provided herein can be applied with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to any plant species susceptible to a plant disease.
  • a plant susceptible to a plant disease is meant that the causative pathogen(s) of the plant disease are able to infect the plant.
  • plant species of interest include, but are not limited to, com ( Zea mays), Brassica sp. (e.g., B. napus, B. rapa, B. juncea), particularly those Brassica species useful as sources of seed oil, alfalfa ( Medicago sativa), rice ( Oryza sativa), rye ( Secale cereale), sorghum ( Sorghum bicolor, Sorghum vulgare), millet (e.g., pearl millet ( Pennisetum glaucum), proso millet ( Panicum miliaceum), foxtail millet ( Setaria italica), finger millet ( Eleusine coracana)), sunflower ( Helianthus annuus), safflower ( Carthamus tinctorius), wheat ( Triticum aestivum), soybean ( Glycine max), tobacco ( Nicotiana tabacum), potato (Solarium tuberosum), peanuts (Arachis hypogaea), cotton ( Gosss
  • Vegetables include tomatoes (Lycopersicon esculentum), lettuce (e.g., Lactuca sativa), green beans (Phaseolus vulgaris), lima beans (Phaseolus limensis), peas (Lathyrus spp.), and members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C. cantalupensis), and musk melon (C. melo).
  • tomatoes Locopersicon esculentum
  • lettuce e.g., Lactuca sativa
  • green beans Phaseolus vulgaris
  • lima beans Phaseolus limensis
  • peas Lathyrus spp.
  • members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C. cantalupensis), and musk melon (C. melo).
  • Ornamentals include azalea (Rhododendron spp.), hydrangea (Macrophylla hydrangea), hibiscus (Hibiscus rosasanensis), roses (Rosa spp.), tulips (Tulipa spp.), daffodils (Narcissus spp.), petunias (Petunia hybrida), carnation (Dianthus caryophyllus), poinsettia (Euphorbia pulcherrima), and chrysanthemum.
  • Conifers that may be employed in practicing the present invention include, for example, pines such as loblolly pine (Pinus taeda), slash pine (Pinus elliotii), ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), and Monterey pine (Pinus radiata); Douglas-fir (Pseudotsuga menziesii), Western hemlock (Tsuga canadensis), Sitka spruce (Picea glauca); redwood (Sequoia sempervirens); true firs such as silver fir (Abies amabilis) and balsam fir (Abies balsamea); and cedars such as Western red cedar (Thuja plicata) and Alaska yellow-cedar (Chamaecyparis nootkatensis).
  • pines such as loblolly pine (Pinus taeda), slash pine (P
  • plants treated with the present combinations and methods are crop plants (for example, com, alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut, sorghum, wheat, millet, tobacco, etc.).
  • plants treated with the present combinations and methods are com (maize), sorghum, wheat, sunflower, tomato, crucifers, peppers, potato, cotton, rice, soybean, sugarbeet, sugarcane, tobacco, barley, oilseed rape, Brassica sp., alfalfa, rye, millet, safflower, peanuts, sweet potato, cassava, coffee, coconut, pineapple, citms trees, cocoa, tea, banana, avocado, fig, guava, mango, olive, papaya, cashew, macadamia, almond, oats, vegetables, ornaments, and conifers.
  • com and soybean plants are optimal, and in yet other embodiments com plants are optimal.
  • plants of interest include grain plants that provide seeds of interest, oil-seed plants, and leguminous plants.
  • Seeds of interest include grain seeds, such as com, wheat, barley, rice, sorghum, rye, etc.
  • Oil-seed plants include cotton, soybean, safflower, sunflower, Brassica, maize, alfalfa, palm, coconut, etc.
  • Leguminous plants include beans, peas, and dry pulses. Beans include guar, locust bean, fenugreek, soybean, garden beans, cowpea, mungbean, lima bean, fava bean, lentils, chickpea, etc.
  • the bacterial strain or active variant thereof provided herein can be applied (simultaneously or sequentially) with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to grapes (bloom to harvest), asparagus, stone fruits (bloom to harvest), and small fruits such as strawberry, blueberry, caneberry.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to grapes (bloom to harvest), asparagus, stone fruits (bloom to harvest), and small fruits such as strawberry, blueberry, caneberry.
  • the bacterial strain and/or the at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole are applied (simultaneously or sequentially) at less than the suggested amount of the bacterial strain and the synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole when applied alone.
  • the bacterial strain can be applied at about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or about 20-70%, about 30-60%, about 30-70%, about 40-80%, about 40-70%, about 40-60%, about 40-50%, about 50-90%, about 50-80%, about 50-70%, about 50-60%, about 60-90%, about 60-80%, about 60-70%, about 70-90%, about 70-90%, or about 80-90% of the suggested amount for application when used alone (i.e., not in conjunction with an additional fungicide).
  • the suggested amount for application of the bacterial strain can be about 1 lb/acre, about 1.25 lb/acre, about 1.5 lb/acre, about 1.75 lb/acre, about 2 lb/acre, about 2.25 lb/acre, about 2.5 lb/acre, about 2.75 lb/acre, about 3 lb/acre, about 3.5 lb/acre, about 4 lb/acre, about 5 lb/acre, about 6 lb/acre, about 7 lb/acre, about 8 lb/acre, about 1-10 lb/acre, or about 1-8 lb/acre.
  • the suggested amount for application of bacterial strain AIP1620, or an active variant thereof can be about 2.5 lb/acre.
  • a formulated product comprising AIP1620 may have 50% AIP1620 by weight, where AIP1620 comprises at least about 400pg of pyrrolnitrin per gram of cells.
  • the application rate of the synthetic fungicide disclosed herein can be calculated based on the application rate of the bacterial strain, or active varian thereof, according to the selected active ingredient weight ratio as disclosed elsewhere herein for each selected synthetic fungicide.
  • Application of 5 lbs/acre of the 50% formulated product comprises 2.5 lbs/acre of the bacterial strain AIP1620.
  • the synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole can be applied at about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or about 20-70%, about 30-60%, about 30-70%, about 40-80%, about 40-70%, about 40-60%, about 40-50%, about 50-90%, about 50-80%, about 50-70%, about 50-60%, about 60-90%, about 60-80%, about 60-70%, about 70-90%, about 70-90%, or about 80-90% of the suggested amount for application when used alone.
  • plant diseases which can be treated or reduced or prevented include, but are not limited to, plant diseases caused by fungi, viruses or viroids, bacteria, insects, nematodes, protozoa, and the like.
  • fungal plant diseases include, but are not limited to, Asian Soybean Rust (ASR), gray mold, leaf spot, Frogeye Leaf Spot, Early Blight, Damping off complex, Brown Patch, black scurf, root rot, belly rot, sheath blight, Powdery Mildew, Anthracnose leaf spot, Downy Mildew, Aerial Blight, Botrytis Blight, Dollar Spot, Fusarium Patch, Pythium Root Rot, Pythium Crown Rot, Stem and Root Rot, Pythium Blight, Late Blight, Fusarium Head Blight, sudden death syndrome (SDS), Fusarium Wilt, Com Stalk Rot, Brown Rust, Black Rust, Yellow Rust, Wheat Rust, Rust, Apple Scab, Verticillium Wil
  • the bacterial strain or active variant thereof provided herein can be combined (in a composition or applied simultaneously or sequentially) with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to target monilinia, botrytis, powdery mildew, phytophthora, rhizoctonia, corynespora,retemaria, sclerotinia, bremia, pseudoperonospora, podosphaera, gleocercospora, and/or fusarium.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole to target monilinia, botrytis,
  • Plant pathogens of the invention include, but are not limited to, viruses or viroids, bacteria, insects, nematodes, fungi, and the like.
  • the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole with the various bacterial strains provided herein target one or more fungal or fungal -like pathogens that cause plant disease.
  • the combination (applied simultaneously or sequentially) of the synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole
  • the bacterial strains provided herein or active variants thereof can have antifungal activity against one, two, three, four, five, or more fungal or fungal-like pathogens and/or fungal or fungal-like diseases described herein.
  • a fungal pathogen or fungal-like pathogen can be, but is not limited to, a fungus or fungal-like organism selected from the group consisting of Botrytis spp., Botrytis cinerea, Cersospora spp., Cercospora sojina, Cercospora beticola, Corynespora spp., Corynespora cassiicola, Altemaria spp., Altemaria dauci, Altemaria solani, Blumeria graminis, Rhizoctonia spp., Rhizoctonia solani, Blumeria graminis f.
  • Colletotrichum gloeosporiodes Discula fraxinea, Gleocercospora spp., Gleocercospora sorghi, Leveillula taurica, Mycosphaerella spp., Phomopsis spp., Plasmopara viticola, Pseudoperonospora spp.,
  • the fungal pathogen or fungal-like pathogen is selected from the group consisting of Ascomycota spp.,Botrytis spp., Botrytis cinerea, Blumeria graminis, Bremia lactucae, Corynespora cassiicola, Cercospora sojina, Alternaria dauci, Alternaria solani, Rhizoctonia solani,
  • the fungal pathogen is Phakopsora sp., including Phakopsora pachyrhizi and/or Phakopsora meibomiae.
  • the presently disclosed combinations do not control Phakopsora sp., such as Phakopsora pachyrhizi.
  • the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole with at least one bacterial strain provided herein target one or more insect or insect pests.
  • the term "insects” or“insect pests” as used herein refers to insects and other similar pests such as, for example, those of the order Acari including, but not limited to, mites and ticks.
  • Insect pests of the present invention include, but are not limited to, insects of the order Lepidoptera, e.g.
  • Mamestra configurata Manduca quinquemaculata, Manduca sexta, Maruca testulalis, Melanchra picta, Operophtera brumata, Orgyia sp., Ostrinia nubilalis, Paleacrita vernata, Papilio cresphontes, Pectinophora gossypiella, Phryganidia californica, Phyllonorycter blancardella, Pieris napi, Pieris rapae, Plathypena scabra, Platynota flouendana, Platynota stultana, Platyptilia carduidactyla, Plodia interpunctella, Plutella xylostella, Pontia protodice, Pseudaletia unipuncta, Pseudoplasia includens, Sabulodes aegrotata, Schizura concin
  • Insect pests also include insects selected from the orders Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera, Isoptera, Anoplura,
  • Insect pests of the invention for the major crops include, but are not limited to: Maize: Ostrinia nubilalis, European com borer; Agrotis ipsilon, black cutworm; Helicoverpa zeae, com earworm; Spodoptera frugiperda, fall armyworm; Diatraea grandiosella, southwestern com borer; Elasmopalpus lignosellus, lesser cornstalk borer; Diatraea saccharalis, surgarcane borer; western com rootworm, e.g., Diabrotica virgifera virgifera; northern com rootworm, e.g., Diabrotica longicornis barberi; southern com rootworm, e.g., Diabrotica undecimpunctata howardi; Melanotus spp., wireworms; Cyclocephala borealis,
  • Cyclocephala immaculata southern masked chafer (white gmb); Popillia japonica, Japanese beetle;
  • Chaetocnema pulicaria com flea beetle
  • Sphenophorus maidis maize billbug
  • Rhopalosiphum maidis com leaf aphid
  • Anuraphis maidiradicis com root aphid
  • Blissus leucopterus leucopterus chinch bug
  • Tetranychus urticae Tetranychus urticae, two-spotted spider mite; Wheat: Pseudaletia unipunctata, army worm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; southern com rootworm, e.g., Diabrotica undecimpunctata howardi, Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid;
  • Cylindrocupturus adspersus sunflower stem weevil; Smicronyx fulus, red sunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil; Suleima helianthana, sunflower bud moth; Homoeosoma electellum, sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus, carrot beetle;
  • Helicoverpa zea cotton bollworm; Spodoptera exigua, beet armyworm; Pectinophora gossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis; Aphis gossypii, cotton aphid; Pseudatomoscelis seriatus, cotton fleahopper; Trialeurodes abutilonea, bandedwinged whitefly; Lygus lineolaris, tarnished plant bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differentialis, differential grasshopper; Thrips tabaci, onion thrips; Franklinkiella fusca, tobacco thrips; Tetranychus cinnabarinus , carmine spider mite; Tetranychus urticae, two-spotted spider mite; Rice: Diatraea saccharalis, sugarcane borer; Spodoptera frugiperda, fall
  • Epilachna varivestis Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, green stink bug; Melanoplus femurrubrum, redlegged grasshopper;
  • the methods and compositions provided herein can also be used against Hemiptera such as Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis Popp, Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus,
  • Pests of interest also include Araecerus fasciculatus , coffee bean weevil;
  • Macrodactylus subspinosus rose chafer; Tribolium confusum, confused flour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum, red flour beetle; Tenebrio molitor, yellow mealworm.
  • Nematodes include parasitic nematodes such as root-knot, cyst, and lesion nematodes, including Heterodera spp., Meloidogyne spp., and Globodera spp.; particularly members of the cyst nematodes, including, but not limited to, Heterodera glycines (soybean cyst nematode); Heterodera schachtii (beet cyst nematode); Heterodera avenae (cereal cyst nematode); and Globodera rostochiensis and Globodera pailida (potato cyst nematodes).
  • Lesion nematodes include Pratylenchus spp.
  • Insect pests can be tested for pesticidal activity of compositions of the invention in early developmental stages, e.g., as larvae or other immature forms.
  • the insects may be reared in total darkness at from about 20 degrees C to about 30 degrees C and from about 30% to about 70% relative humidity.
  • Bioassays may be performed as described in Czapla and Lang (1990) J. Econ. Entomol. 83 (6): 2480-2485. Methods of rearing insect larvae and performing bioassays are well known to one of ordinary skill in the art.
  • the bacterial strains or active variants thereof i.e., AIP1620, AIP050999, and CGA267356, or an active variant of any thereof
  • Podosphaera xanthii Golovinomyces cichoracearum, Erysiphe lager stroemiae, Gleocercospora spp., Gleocercospora sorghi, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errahunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea, Plasmopara viticola,
  • Pseudoperonospora spp . Pseudoperonospora cubensis, Peronospora belbahrii, Bremia lactucae,
  • Peronospora lamii Plasmopara obduscens, Pythium cryptoirregulare, Pythium aphanidermatum, Pythium irregulare, Pythium sylvaticum, Pythium myriotylum, Pythium ultimum, Phytophthora capsici, Phytophthora nicotianae, Phytophthora infe stans, Phytophthora tropicalis, Phytophthora sojae, Sclerotinia spp.,
  • Gleocercospora sorghi Rhizoctonia solani, Erysiphe necator, Podosphaera xanthii, Colletotrichum cereal, Plasmopara viticola, Peronospora belbahrii, Pythium aphanidermatum, Pythium sylvaticum, Pythium myriotylum, Pythium ultimum, Phytophthora nicotianae, Phytophthora infestans, Phytophthora tropicalis, Phytophthora sojae, Pseudoperonospora cubensis, Fusarium graminearum, Fusarium solani, Phakopsora pachyrizi, Sclerotinia minor, and Venturia inaequalisa.
  • the bacterial strains or active variants thereof i.e., AIP1620, AIP050999, and CGA267356, or an active variant of any thereof
  • the synthetic fungicide and the bacterial strain or modified biological agents disclosed herein i.e., AIP1620, AIP050999, and CGA267356, or an active variant of any thereof, or a combination of cells from any one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof
  • synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole with the bacterial strains or modified bacterial strains disclosed herein (i.e.
  • a plant disease comprising applying to a plant or plant part having a plant disease or at risk of developing a plant disease an effective amount of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole in combination (applied simultaneously or sequentially) with at least one bacterial strain provided herein or an active variant thereof, wherein the combination controls a plant pathogen that causes the plant disease.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole in combination (applied simultaneously or sequentially) with at least one bacterial strain provided herein or an active variant thereof, wherein the combination controls a plant pathogen that causes the plant disease.
  • the bacterial strain provided herein or active variant thereof may comprise at least one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof, or a combination of cells from any one of AIP1620, AIP050999, and
  • the effective amount of the presently disclosed combination comprises an active ingredient weight ratio of the bacterial strain or active variant thereof to the synthetic fungicide as disclosed herein for the selected synthetic fungicide wherein the combination controls a plant pathogen, treats or prevents a plant disease, or improves an agronomic trait of interest in a plant.
  • the effective amount of the combination can comprise at least about 4.4xl0 5 pg of pyrrolnitrin per acre or least about 1 x 10 4 to 1 x 10 6 pg of pyrrolnitrin per acre of the bacterial strain or active variant thereof.
  • AIP1620 is applied at 2.5 lb/acre wherein the formulation comprises at least about 400pg of pyrrolnitrin per gram of cells.
  • Applying AIP1620, or active variant thereof, in the presence of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole can reduce the amount of synthetic fungicide or bacterial strain required for a given result.
  • the amount of the bacterial strain can be less than 2.5 lb/acre, such as between about 0.1 lb/acre and 2.49 lb/acre, including but not limited to about 2.49 lb/acre, about 2.45 lb/acre, about 2.4 lb/acre, about 2.3 lb/acre, about 2.2 lb/acre, about 2.1 lb/acre, about 2.0 lb/acre, about 1.9 lb/acre, about 1.8 lb/acre, about 1.7 lb/acre, about 1.6 lb/acre, about 1.5 lb/acre, about 1.4 lb/acre, about 1.3
  • the amount of the synthetic fungicide can be reduced by at least about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or more, or about 1-5%, about 5- 10%, about 5-15%, about 5-20%, about 1-10%, about 1-15%, about 1-20%, about 1-25%,
  • the application rate of the synthetic fungicide disclosed herein can be calculated based on the rate of the bacterial strain, or active varian thereof, according to the selected active ingredient weight ratio as disclosed elsewhere herein for each selected synthetic fungicide.
  • the formulated product that is applied that comprises AIP1620 may have about 50% AIP1620 by weight, where AIP1620 comprises at least about 400pg of pyrrolnitrin per gram of cells. Application of 5 lbs/acre of the 50% formulated product comprises 2.5 lbs/acre of the bacterial strain AIP1620.
  • the bacterial strain and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole can be applied (simultaneously or sequentially) to a plant, plant plart, or area of plant cultivation in a weight ratio that results in at least an additive effect when applied to a plant, plant part, or area of plant cultivation.
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with tetraconazole or flutriafol in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial strain: synthetic fungicide) to about 1000: 1, including but not limited to about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about2:l, about3:l, about 4: 1, about 5:1, about 6: 1, about 7:1, about 8: 1, about 9:1, about 10: 1, about 15:1, about 20: 1, about 25:1, about 30:1, about 40: 1, about 50: 1, about 60: 1, about 70: 1, about 80: 1, about 90: 1, about 100: 1, about 150: 1, about 200: 1, about 300: 1, about 400: 1, about 500: 1, about 600: 1, about 700: 1, about 800: 1, about 900:1 or about 1000:1.
  • the bacterial strain, or active variant thereof can be applied
  • tetraconazole or flutriafol in an active ingredient weight ratio (lb/lb) of about 1:1 (bacterial strain: synthetic fungicide) to about 100: 1, including but not limited to about 1:1, about 2:1, about 5:1, about 10:1, about 15:1, about 20:1, about 25:1, about 30:1, about 35:1, about 40:1, about 45:1, about 50:1, about 55:1, about 60:1, about 61:1, about 62:1, about 62.5:1, about 63:1, about 64:1, about 65:1, about 66: 1, about 67:1, about 68: 1, about 69: 1, about 70: 1, about 75:1, about 80: 1, about 85:1, about 90: 1, about 95: 1, and about 100: 1.
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with azoxystrobin, tebuconazole, or difenoconazole in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial strain synthetic fungicide) to about 500: 1, about 1 :5 to about 100:1, about 1:5 to about 75:1, about 1:5 to about 50:1, about 1:5 to about 20:1, about 1:1 to about 100:1, about 1:1 to about 75:1, about 1 : 1 to about 50 : 1 , or about 1 : 1 to about 20:1, including but not limited to about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about2:l, about3:l, about 4: 1, about 5:1, about 6: 1, about 7:1, about 8: 1, about 9:1, about 10: 1, about 15:1, about 20: 1, about 25:1,
  • the bacterial strain, or active variant thereof can be applied
  • azoxystrobin in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain: synthetic fungicide) to about 25:1 or about 1: 1 to about 20: 1, including but not limited to about 1:1, about 1.5:1, about2:l, about2.5:l, about3:l, about3.5:l, about 4:1, about 4.5:1, about5:l, about 5.5:1, about 6:1, about 6.1:1, about 6.2:1, about 6.3:1, about 6.4:1, about 6.5:1, about 6.6:1, about 6.7:1, about 6.8:1, about 6.9:1, about 7:1, about 7.1:1, about 7.2:1, about 7.3:1, about 7.4:1, about 7.5:1, about 7.6:1, about 7.7:1, about 7.8:1, about 7.9:1, about 8:1, about 8.5:1, about 9:1, about 9.5:1, about 10:1, about 10.25:1, about 10.4:1, about 10.5:1, about 10.75:1, about 11:
  • the bacterial strain, or active variant thereof can be applied
  • tebuconazole in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain: synthetic fungicide) to about 50:1, including but not limited to about 1:1, about 3:1, about 5:1, about 7:1, about 9:1, about 11:1, about 13:1, about 15:1, about 17:1, about 17.5:1, about 18:1, about 18.5:1, about 19:1, about 19.5:1, about 20:1, about 16:1, about 17:1, about 18:1, about 19:1, about 19.1:1, about 19.2:1, about 19.3:1, about 19.4:1, about 19.5:1, about 19.6:1, about 19.7:1, about 19.8:1, about 19.9:1, about 20:1, about 20.25:1, about 20.5:1, about 20.75:1, about21:l, about21.25:l, about21.5:l, about 21.75:1, about 22:1, about 22.2:1, about 22.22:1, about 22.25:1, about 22.5:1, about 22.75
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with difenconazole in an active ingredient weight ratio (lb/lb) of about 1:1 (bacterial strain synthetic fungicide) to about 50: 1, about 1 : 1 to about 40: 1, about 10: 1 to about 40: 1, or about 10: 1 to about 30:1, including but not limited to, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26:1, about 27:1, about 28:1, about 29:1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35.1:1, about 36:1, about 37:1, about 38:1, about 39:1,
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with chlorothalonil in an active ingredient weight ration (lb/lb) of about 1 : 100 (bacterial strain synthetic fungicide) to about 500: 1, including but not limited to, about 1 : 100, about 1:50, about 1:10, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1.5, about 1:1.3, about 1:1.2, about 1:1.1, about 1:1, about 1.1:1, about 1.12:1, about 1.125:1, about 1.13:1, about 1.15:1, about 1.2:1, about 1.3:1, about 1.4:1, about 1.5:1, about 1.8:1, about 2:1, about2.5:l, about3:l, about 4:1, about5:l, about 10:1, about 50:1, about 100:1, about 150:1, about 200:1, about 250:1, about 300:1, about 350:1, about 400:1, about 450:1, and about 500:1.
  • active ingredient weight ration lb/lb
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with chlorothalonil in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial strain: synthetic fungicide) to about 10:1, including but not limited to, about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1.9, about 1:1.8, about 1:1.75, about 1:1.7, about 1:1.65, about 1:1.6, about 1:1.55, about 1:1.5, about 1:1.45, about 1:4, about 1:1.35, about 1:1.3, about 1:1.275, about 1:1.25, about 1:1.24, about 1:1.23, about 1:1.22, about 1:1.21, about 1:1.2, about 1:1.19, about 1:1.18, about 1:1.17, about 1:1.16, about 1:1.15, about 1:1.15, about 1:1.1, about 1:1, about 1.1:1, about 1.2:1, about 1.3:1, about 1.4:
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with triflumizole in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial straimtriflumizole) to about 100: 1, including but not limited to, about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 2:1, about3:l, about 4:1, about5:l, about 6: 1, about 7:1, about 8:1, about 9:1, about 10: 1, about 15:1, about 20: 1, about 25:1, about 30: 1, about 35:1, about 40:1, about 45:1, about 50:1, about 55:1, about 60:1, about 65:1, about 70:1, about 75:1, about 80:1, about 85:1, about 90:1, about 95:1, or about 100:1
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with triflumizole in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial straimtriflumizole) to about 25:1 or about 1:10 to about 20:1, including but not limited to about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 1.1:1, about 1.2:1, about 1.3:1, about 1.4:1, about 1.5:1, about 1.6:1, about 1.66:1, about 1.7:1, about 1.8:1, about 1.9:1, about 2:1, about 2:1, about 2.5:1, about 3:1, about 3.5:1, about 4:1, about 4.5:1, about 5:1, about 5.5:1, about 6:1, about 6.5:1, about 7:1, about 7.5:1, about 8:1, about 8.5:1, about 9:1, about 9.5:1, about 10:1, about 15:1, about 20:1,
  • the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole with a bacterial strain provided herein or an active variant thereof treats or prevents one, two, three, four, five or more plant diseases.
  • the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole with a bacterial strain provided herein or an active variant thereof treats or prevents one, two, three, four, five or more fungal and/or fungal-like plant diseases.
  • the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole with a bacterial strain provided herein or an active variant thereof can be employed with any plant species susceptible to a plant disease of interest.
  • Table 1 Examples of diseases causes by the fungal or fungal-like pathogens described herein are provided in Table 1. Also provided are non-limiting exemplary crop species that are susceptible to the plant diseases caused by the pathogens. For example, Table 1 shows that Bortrytis cinerea causes gray mold on all flowering crops.
  • a combination of a bacterial strain provided herein or active variant thereof and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole that controls Bortrytis cinerea can be applied (simultaneously or sequentially) to a plant having gray mold or at risk of developing gray mold in order to treat or prevent gray mold in the plant.
  • Table 1 shows that Rhizoctonia solani causes Damping off complex in com, Damping off complex in soybean, Brown Patch in turf, and Damping off complex in ornamentals.
  • a combination of a bacterial strain provided herein or active variant thereof and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole that controls Rhizoctonia solani can be applied (simultaneously or sequentially) to a plant having Damping off complex and/or brown patch or at risk of developing Damping off complex and/or brown patch in order to treat or prevent Damping off complex and/or brown patch in the plant.
  • Table 1 shows that Colletotrichum cereal, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea cause Anthracnose leaf spot.
  • a combination of a bacterial strain provided herein or active variant thereof and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole that controls one or more of Colletotrichum cereal, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea can be applied (simultaneously or sequentially) to a plant having Anthracnose leaf spot or at risk of developing Anthracnose leaf spot in order to treat or prevent Anthracnose leaf spot in the plant.
  • ASR Asian Soybean Rust
  • methods of treating or preventing Asian Soybean Rust comprising applying to a plant having ASR or at risk of developing ASR an effective amount of a combination (applied simultaneously or sequentially) of at least one bacterial strain provided herein or an active variant thereof comprising AIP1620, AIP050999, and CGA267356, or an active variant of any thereof; or a combination of cells from any one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof along with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • the effective amount comprises applying the combination of the bacterial strain and synthetic fungicide at a weight ratio disclosed herein for the selected synthetic fungicide, wherein the combination comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells, wherein the bacterial strain provided herein or active variant thereof in combination with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole controls a plant pathogen that causes ASR.
  • chlorothalonil, and difenoconazole and at least one bacterial strain provided herein or active variant thereof provided herein is used as a foliar application on a plant to treat or prevent ASR.
  • the application rate of the synthetic fungicide disclosed herein can be calculated based on the rate of the bacterial strain, or active varian thereof, according to the selected active ingredient weight ratio as disclosed elsewhere herein for each selected synthetic fungicide.
  • chlorothalonil, and difenoconazole with a bacterial strain provided herein or an active variant thereof or modified bacterial strain provided herein can be employed with any plant species susceptible to ASR.
  • a plant susceptible to Asian Soybean Rust (ASR) is meant that the causative pathogen(s) of ASR are able to infect the plant.
  • soybean ' Glycine max
  • common bean Phaseolus vulgaris
  • lima beans Phaseolus limensis
  • cowpeas Vigna unguiculata
  • pigeon peas Cajanus cajan
  • yam beans such as jicama ( Pachyrhizus erosus).
  • a soybean plant is employed.
  • ASR is caused by one or more fungal pathogens of the genus Phakopsora.
  • the fungal pathogens that cause ASR are Phakopsora pachyrhizi or Phakopsora meibomiae.
  • the ASR pathogen is well adapted for long-distance dispersal, because the spores can be readily carried by the wind, making it an ideal means for introduction to new, rust-free regions.
  • the primary means of dissemination are spores, which can be carried by wind or splashed rain. These pathogens are obligate pathogens surviving and reproducing only on live hosts.
  • the first symptoms are light-brown polygonal lesions of 2 to 5 mm on the adaxial leaf surface. These lesions develop into volcano-shaped lesions known as pustules that appear on the abaxial surface of the leaf, where uredospores are produced.
  • a combination (applied simultaneously or sequentially) of a bacterial strain provided herein or an active variant thereof and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole controls Phakopsora pachyrhizi.
  • a combination (applied simultaneously or sequentially) of a bacterial strain provided herein or active variant thereof and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole controls Phakopsora meihomiae.
  • synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole controls Phakopsora meihomiae.
  • synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole controls Phakopsora meih
  • “treat” or“treating” or its derivatives includes substantially inhibiting, slowing, or reversing the progression of a condition, substantially ameliorating symptoms of a condition or substantially preventing the appearance of symptoms or conditions brought about by the pathogen that causes the plant disease.
  • controlling and“protecting a plant from a pathogen” refers to one or more of inhibiting or reducing the growth, germination, reproduction, and/or proliferation of a pathogen of interest; and/or killing, removing, destroying, or otherwise diminishing the occurrence, and/or activity of a pathogen of interest.
  • a plant treated with the bacterial strain provided herein and synthetic fungicide may show a reduced disease severity or reduced disease development in the presence of plant pathogens by a statistically significant amount.
  • prevent and its variations means the countering in advance of bacterial, fungal, viral, insect or other pest growth, proliferation, infestation, spore germination, and hyphae growth.
  • the composition is applied before exposure to the pathogens.
  • ameliorate and“amelioration” relate to the improvement in the treated plant condition brought about by the compositions and methods provided herein.
  • the improvement can be manifested in the forms of a decrease in pathogen growth and/or an improvement in the diseased plant height, weight, number of leaves, root system, or yield.
  • the term refers to the improvement in a diseased plant physiological state.
  • inhibitor and all variations of this term is intended to encompass the restriction or prohibition of bacterial, fungal, viral, nematode, insect, or any other pest growth, as well as spore germination.
  • eliminate relates to the substantial eradication or removal of bacteria, fungi, viruses, nematodes, insects, or any other pests by contacting them with the composition of the invention, optionally, according to the methods of the invention described below.
  • delay means the slowing of the progress of bacterial, fungal, viral, nematode, insect, or any other pest growth, and spore germination.
  • the expression “delaying the onset” is interpreted as preventing or slowing the progression of bacterial, fungal, viral, nematodes, insect, or any other pest growth, infestation, infection, spore germination and hyphae growth for a period of time, such that said bacterial, fungal, viral, nematode, insect, or any other pest growth, infestation, infection, spore germination and hyphae growth do not progress as far along in development, or appear later than in the absence of the treatment according to the invention.
  • a plant, plant part, or area of cultivation treated with a combination (applied simultaneously or sequentially) of a bacterial strain provided herein or an active variant thereof and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole may show a reduced disease severity or reduced disease development in the presence of plant pathogens by a statistically significant amount.
  • a reduced disease severity or reduced disease development can be a reduction of about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% when compared to non- treated control plants.
  • the plant treated with a combination (applied simultaneously or sequentially) of a bacterial strain provided herein or an active variant thereof and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole may show a reduced disease severity or reduced disease development in the presence of plant pathogen at least about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%,
  • Methods for assessing plant disease severity include, measuring percentage of diseased leaf area (Godoy et al. (2006) Fitopatol. Bras. 31(1) 63-68) or by measuring uredinia counts (see Example 1).
  • synergy or“synergistically” is intended that the combination (applied simultaneously or sequentially) of a bacterial strain provided herein, or an active variant thereof, and a synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole can (in some embodiments) provide a greater control of plant disease or a plant pathogen and/or improvement of at least one agronomic trait of interest than the additive effect of the bacterial strain and the synthetic fungicide applied individually (i.e., in the absence of the other).
  • This synergy can be an increase of 2%, 5%, 7%, 10%, 12%, 15%, 17%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 100%, or more above the additive effect of the application of the bacterial strain and the synthetic fungicide alone (i.e., in the absence of the other).
  • the application (simultaneous or sequential) of a bacterial strain provided herein, or an active variant thereof, and a synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole function synergistically when no control of a plant pathogen or treatment or prevention of a plant disease or improvement of at least one agronomic trait of interest is exhibited when the bacterial strain or synthetic fungicide is used alone, but the application (simultaneous or sequential) of the two results in control of a plant pathogen or treatment or prevention of a plant disease or improvement of at least one agronomic trait of interest.
  • an antipathogenic composition reduces the disease symptoms resulting from pathogen challenge by a statistically significant amount, including for example, at least about 10% to at least about 20%, at least about 20% to about 50%, at least about 10% to about 60%, at least about 30% to about 70%, at least about 40% to about 80%, or at least about 50% to about 90% or greater.
  • the methods of the invention can be utilized to protect plants from disease, particularly those diseases that are caused by plant pathogens.
  • Assays that measure antipathogenic activity are commonly known in the art, as are methods to quantitate disease resistance in plants following pathogen infection. See, for example, U.S. Patent No. 5,614,395, herein incorporated by reference. Such techniques include, measuring overtime, the average lesion diameter, the pathogen biomass, and the overall percentage of decayed plant tissues. For example, a plant having an antipathogenic composition applied to its surface shows a decrease in tissue necrosis (i.e., lesion diameter) or a decrease in plant death following pathogen challenge when compared to a control plant that was not exposed to the antipathogenic composition. Alternatively, antipathogenic activity can be measured by a decrease in pathogen biomass. For example, a plant exposed to an antipathogenic composition is challenged with a pathogen of interest.
  • RNA samples from the pathogen- inoculated tissues are obtained and RNA is extracted.
  • the percent of a specific pathogen RNA transcript relative to the level of a plant specific transcript allows the level of pathogen biomass to be determined. See, for example, Thomma et al. (1998) Plant Biology 95: 15107-15111, herein incorporated by reference.
  • in vitro antipathogenic assays include, for example, the addition of varying concentrations of the antipathogenic composition to paper disks and placing the disks on agar containing a suspension of the pathogen of interest. Following incubation, clear inhibition zones develop around the discs that contain an effective concentration of the antipathogenic composition (Liu et al. (1994) Plant Biology 91: 1888-1892, herein incorporated by reference). Additionally, microspectrophotometrical analysis can be used to measure the in vitro antipathogenic properties of a composition (Hu et al. (1997) Plant Mol. Biol. 34:949-959 and Cammue et al. (1992) J. Biol. Chem. 267: 2228-2233, both of which are herein incorporated by reference).
  • compositions and methods for inducing disease resistance in a plant to plant pathogens are also provided. Accordingly, the compositions and methods are also useful in protecting plants against fungal and/or fungal -like pathogens, viruses, nematodes, and insects.
  • bacterial strain provided herein or active variant thereof may comprise at least one of AIP1620, AIP050999, and
  • the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole with the bacterial strain provided herein or active variant thereof promotes at least an additive defensive response to the pathogen that causes the plant disease.
  • the effective amount of the combination comprises a bacterial strain provided herein or active variant thereof in combination with a synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole in a weight ratio (lb/lb) disclosed herein for the selected synthetic fungicide.
  • Hie application rate of the synthetic fungicide disclosed herein can be calculated based on the rate of the bacterial strain, or active varian thereof, according to the selected active ingredient weight ratio as disclosed elsewhere herein for each selected synthetic fungicide.
  • the combination comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells.
  • a defensive response in the plant can be triggered after applying the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and a bacterial strain provided herein or active variant thereof to the plant, but prior to pathogen challenge and/or after pathogen challenge of the plant treated with the combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole,
  • the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole with a bacterial strain provided herein or active variant thereof induces resistance to one, two, three, four, five or more plant pathogens described herein.
  • the bacterial strain provided herein or active variant thereof and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole induces resistance to one, two, three, four, five or more fungal and/or fungal-like plant pathogens described herein.
  • disease resistance is intended that the plants avoid the disease symptoms that result from plant- pathogen interactions. That is, pathogens are prevented from causing plant diseases and the associated disease symptoms, or alternatively, the disease symptoms caused by the pathogen are minimized or lessened as compared to a control.
  • methods of improving plant health and/or improving an agronomic trait of interest comprising applying to a plant an effective amount of a combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain provided herein or an active variant thereof or an active derivative thereof.
  • the bacterial strain provided herein or active variant thereof may comprise at least one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof; or a combination of cells from any one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof.
  • the effective amount of the combination comprises a bacterial strain provided herein or active variant thereof in combination with a synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole,
  • the combination comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells.
  • Increased plant health is meant increased growth and/or yield of a plant, increased stress tolerance and/or decreased herbicide resistance, to name a few.
  • Increased stress tolerance refers to an increase in the ability of a plant to decrease or prevent symptoms associated with one or more stresses.
  • the stress can be a biotic stress that occurs as a result of damage done to plants by other living organisms such as a pathogen (for example, bacteria, viruses, fungi, parasites), insects, nematodes, weeds, cultivated or native plants.
  • the stress can also be an abiotic stress such as extreme temperatures (high or low), high winds, drought, salinity, chemical toxicity, oxidative stress, flood, tornadoes, wildfires, radiation and exposure to heavy metals.
  • an effective amount of the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutrafiol, tebuconazole, chlorothalonil, and difenoconazole, and the bacterial strain or active variant thereof improves plant health or improves an agronomic trait of interest by at least an additive amount.
  • an effective amount of the combination (applied simultaneously or sequentially) of a synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and the bacterial strain or active variant thereof improves plant health or improves an agronomic trait of interest by a synergistic amount, including for example, at least about 10% to at least about 20%, at least about 20% to about 50%, at least about 10% to about 60%, at least about 30% to about 70%, at least about 40% to about 80%, or at least about 50% to about 90% or greater than the additive effect of the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) and the bacterial strain applied alone.
  • a synthetic fungicide
  • a bacterial strain provided herein or active variant thereof and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole is applied (simultaneously or sequentially) in an effective amount.
  • the effective amount of the combination comprises an amount of the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) and/or the bacterial strain provided herein that is lower than the suggested use or standard use in the art.
  • the synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole
  • An effective amount of a combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and bacterial strain provided herein or active variant thereof is an amount sufficient to control, treat, prevent, inhibit the pathogen that causes a plant disease, and/or reduce plant disease severity or reduce plant disease development when applied at an application rate lower than the suggested use or standard use.
  • the effective amount of a combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and a bacterial strain provided herein or active variant thereof is an amount sufficient to improve an agronomic trait of interest and/or to promote or increase plant health, growth or yield of a plant susceptible to a disease when used at an effective amount lower than the suggested use or standard use.
  • the rate of application or amount of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and/or the bacterial strain provided herein or active variant thereof may vary according to the pathogen being targeted, the crop or plant to be protected, the efficacy of the bacterial strain provided herein or active variant thereof, the severity of the disease, the climate conditions, the agronomic trait of interest to improve, and the like.
  • the standard or suggested use of bacterial strain AIP1620 is about 2.5 lb/acre wherein the composition.
  • the combination comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells, or about 1 8xl0 5 pg pyrrolnitrin per pound of bacterial cells.
  • the application amount of AIP1620, or an active variant thereof can be less than 2.5 lb/acre, such as between about 0.1 lb/acre and 2.49 lb/acre, including but not limited to about 2.49 lb/acre, about 2.45 lb/acre, about 2.4 lb/acre, about 2.3 lb/acre, about 2.2 lb/acre, about 2.1 lb/acre, about 2.0 lb/acre, about 1.9 lb/acre, about
  • the application rate of the synthetic fungicide disclosed herein can be calculated based on the rate of the bacterial strain, or active varian thereof, according to the selected active ingredient weight ratio as disclosed elsewhere herein for each selected synthetic fungicide.
  • the combination of a bacterial strain provided herein or active variant thereof and tetraconazole is applied (simultaneously or sequentially) in an effective amount, wherein the standard or suggested use of bacterial strain AIP1620 is about 2.5 lb/acre in the combination.
  • the application amount of AIP1620, or an active variant thereof can be less than 8 lb/acre, less than 7 lb/acre, less than 6 lb/acre, less than 5 lb/acre, less than 4 lb/acre, 3 lb/acre, or less than 2.5 lb/acre, such as between about 0.1 lb/acre and 2.49 lb/acre, including but not limited to about 2.49 lb/acre, about 2.45 lb/acre, about 2.4 lb/acre, about 2.3 lb/acre, about
  • the combination of a bacterial strain provided herein or active variant thereof and tetraconazole comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells, or about 1.8xl0 5 pg pyrrolnitrin per pound of bacterial cells.
  • the combination of a bacterial strain provided herein or active variant thereof and triflumizole is applied (simultaneously or sequentially) in an effective amount, wherein the standard or suggested use of bacterial strain AIP1620 is about 2.5 lb/acre in the combination.
  • the application amount of AIP1620, or an active variant thereof can be less than 8 lb/acre, less than 7 lb/acre, less than 6 lb/acre, less than 5 lb/acre, less than 4 lb/acre, 3 lb/acre, or less than 2.5 lb/acre, such as between about 0.1 lb/acre and 2.49 lb/acre, including but not limited to about 2.49 lb/acre, about 2.45 lb/acre, about 2.4 lb/acre, about 2.3 lb/acre, about 2.2 lb/acre, about 2.1 lb/acre, about 2.0 lb/acre, about 1.9 lb/acre, about 1.8 lb/acre, about 1.7 lb/acre, about
  • the combination of a bacterial strain provided herein or active variant thereof and triflumizole comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells, or about 1.8xl0 5 pg pyrrolnitrin per pound of bacterial cells.
  • the combination of a bacterial strain provided herein or active variant thereof and azoxystrobin is applied (simultaneously or sequentially) in an effective amount, wherein the standard or suggested use of bacterial strain AIP1620 is about 2.5 lb/acre in the combination.
  • the application amount of AIP1620, or an active variant thereof can be less than 8 lb/acre, less than 7 lb/acre, less than 6 lb/acre, less than 5 lb/acre, less than 4 lb/acre, 3 lb/acre, or less than 2.5 lb/acre, such as between about 0.1 lb/acre and 2.49 lb/acre, including but not limited to about 2.49 lb/acre, about 2.45 lb/acre, about 2.4 lb/acre, about 2.3 lb/acre, about 2.2 lb/acre, about 2.1 lb/acre, about 2.0 lb/acre, about 1.9 lb/acre, about 1.8 lb/acre, about 1.7 lb/acre, about
  • the combination of a bacterial strain provided herein or active variant thereof and azoxystrobin comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells, or about 1.8xl0 5 pg pyrrolnitrin per pound of bacterial cells.
  • the combination of a bacterial strain provided herein or active variant thereof and flutriafol is applied (simultaneously or sequentially) in an effective amount, wherein the standard or suggested use of bacterial strain AIP1620 is about 2.5 lb/acre in the combination.
  • the application amount of AIP1620, or an active variant thereof can be less than 8 lb/acre, less than 7 lb/acre, less than 6 lb/acre, less than 5 lb/acre, less than 4 lb/acre, 3 lb/acre, or less than 2.5 lb/acre, such as between about 0.1 lb/acre and 2.49 lb/acre, including but not limited to about 2.49 lb/acre, about 2.45 lb/acre, about 2.4 lb/acre, about 2.3 lb/acre, about 2.2 lb/acre, about 2.1 lb/acre, about 2.0 lb/acre, about 1.9 lb/acre, about 1.8 lb/acre, about 1.7 lb/acre, about
  • the combination of a bacterial strain provided herein or active variant thereof and flutriafol comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells, or about 1.8xl0 5 pg pyrrolnitrin per pound of bacterial cells.
  • the combination of a bacterial strain provided herein or active variant thereof and tebuconazole is applied (simultaneously or sequentially) in an effective amount, wherein the standard or suggested use of bacterial strain AIP1620 is about 2.5 lb/acre in the combination.
  • the application amount of AIP1620, or an active variant thereof can be less than 8 lb/acre, less than 7 lb/acre, less than 6 lb/acre, less than 5 lb/acre, less than 4 lb/acre, 3 lb/acre, or less than 2.5 lb/acre, such as between about 0.1 lb/acre and 2.49 lb/acre, including but not limited to about 2.49 lb/acre, about 2.45 lb/acre, about 2.4 lb/acre, about 2.3 lb/acre, about
  • the combination of a bacterial strain provided herein or active variant thereof and tebuconazole comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells, or about 1.8xl0 5 pg pyrrolnitrin per pound of bacterial cells.
  • the combination of a bacterial strain provided herein or active variant thereof and chlorothalonil is applied (simultaneously or sequentially) in an effective amount, wherein the standard or suggested use of bacterial strain AIP1620 is about 2.5 lb/acre in the combination.
  • the application amount of AIP1620, or an active variant thereof can be less than 8 lb/acre, less than 7 lb/acre, less than 6 lb/acre, less than 5 lb/acre, less than 4 lb/acre, 3 lb/acre, or less than 2.5 lb/acre, such as between about 0.1 lb/acre and 2.49 lb/acre, including but not limited to about 2.49 lb/acre, about 2.45 lb/acre, about 2.4 lb/acre, about 2.3 lb/acre, about
  • the combination of a bacterial strain provided herein or active variant thereof and chlorothalonil comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells, or about 1.8xl0 5 pg pyrrolnitrin per pound of bacterial cells.
  • the combination of a bacterial strain provided herein or active variant thereof and difenoconazole is applied (simultaneously or sequentially) in an effective amount, wherein the standard or suggested use of bacterial strain AIP1620 is about 2.5 lb/acre in the combination.
  • the application amount of AIP1620, or an active variant thereof can be less than 8 lb/acre, less than 7 lb/acre, less than 6 lb/acre, less than 5 lb/acre, less than 4 lb/acre, 3 lb/acre, or less than 2.5 lb/acre, such as between about 0.1 lb/acre and 2.49 lb/acre, including but not limited to about 2.49 lb/acre, about 2.45 lb/acre, about 2.4 lb/acre, about 2.3 lb/acre, about 2.2 lb/acre, about 2.1 lb/acre, about 2.0 lb/acre, about 1.9 lb/acre, about 1.8 lb/acre, about 1.7 lb/acre, about
  • the combination of a bacterial strain provided herein or active variant thereof and difenoconazole comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells, or about 1.8xl0 5 pg pyrrolnitrin per pound of bacterial cells.
  • the bacterial strain and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole can be applied (simultaneously or sequentially) as a field inoculation in a weight ratio that results in at least an additive effect.
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with tetraconazole or flutriafol in an active ingredient weight ratio (lb/lb) of about 1 : 10 (bacterial strain: synthetic fungicide) to about 1000: 1, including but not limited to about 1 : 10, about 1 :9, about 1:8, about 1 :7, about 1 :6, about 1 :5, about 1 :4, about 1 :3, about 1 :2, about 1 : 1, about 2: l, about 3: l, about 4: 1, about 5: 1, about 6: 1, about 7: 1, about 8: 1, about 9: 1, about 10: 1, about 15: 1, about 20: 1, about 25: 1, about 30: 1, about 40: 1, about 50: 1, about 60: 1, about 70: 1, about 80: 1, about 90: 1, about 100: 1, about 150: 1, about 200: 1, about 300: 1, about 400: 1, about 500: 1, about 600: 1, about 700: 1, about 800: 1, about 900: 1 or about
  • the bacterial strain, or active variant thereof can be applied
  • tetraconazole or flutriafol in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain: synthetic fungicide) to about 100: 1, including but not limited to about 1 : 1, about 2: 1, about 5: 1, about 10: 1, about 15: 1, about 20: 1, about 25: 1, about 30: 1, about 35: 1, about 40: 1, about 45: 1, about 50: 1, about 55: 1, about 60: 1, about 61 : 1, about 62: 1, about 62.5: 1, about 63: 1, about 64: 1, about 65: 1, about 66: 1, about 67: 1, about 68: 1, about 69: 1, about 70: 1, about 75: 1, about 80: 1, about 85: 1, about 90: 1, about 95: 1, and about 100: 1.
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with azoxystrobin, tebuconazole, or difenoconazole in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial strain: synthetic fungicide) to about 500: 1, about 1 :5 to about 100:1, about 1:5 to about 75:1, about 1:5 to about 50:1, about 1:5 to about 20:1, about 1:1 to about 100:1, about 1:1 to about 75:1, about 1 : 1 to about 50 : 1 , or about 1 : 1 to about 20:1, including but not limited to about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about2:l, about3:l, about 4: 1, about 5:1, about 6: 1, about 7:1, about 8: 1, about 9: 1, about 10: 1, about 15:1, about 20: 1, about 25:
  • the bacterial strain, or active variant thereof can be applied
  • azoxystrobin in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain: synthetic fungicide) to about 25:1 or about 1: 1 to about 20: 1, including but not limited to about 1:1, about 1.5:1, about2:l, about2.5:l, about3:l, about3.5:l, about 4:1, about 4.5:1, about5:l, about 5.5:1, about 6:1, about 6.1:1, about 6.2:1, about 6.3:1, about 6.4:1, about 6.5:1, about 6.6:1, about 6.7:1, about 6.8:1, about 6.9:1, about 7:1, about 7.1:1, about 7.2:1, about 7.3:1, about 7.4:1, about 7.5:1, about 7.6:1, about 7.7:1, about 7.8:1, about 7.9:1, about 8:1, about 8.5:1, about 9:1, about 9.5:1, about 10:1, about 10.5:1, about 11:1, about 11.5:1, about 12:1, about 12.5:1,
  • the bacterial strain, or active variant thereof can be applied
  • tebuconazole in an active ingredient weight ratio (lb/lb) of about 1 : 1 (bacterial strain: synthetic fungicide) to about 50:1, including but not limited to about 1:1, about 3:1, about 5:1, about 7:1, about 9:1, about 11:1, about 13:1, about 15:1, about 17:1, about 17.5:1, about 18:1, about 18.5:1, about 19:1, about 19.5:1, about 20:1, about 16:1, about 17:1, about 18:1, about 19:1, about 19.1:1, about 19.2:1, about 19.3:1, about 19.4:1, about 19.5:1, about 19.6:1, about 19.7:1, about 19.8:1, about 19.9:1, about 20:1, about 20.25:1, about 20.5:1, about 20.75:1, about21:l, about21.25:l, about21.5:l, about 21.75:1, about 22:1, about 22.2:1, about 22.22:1, about 22.25:1, about 22.5:1, about 22.75
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with difenconazole in an active ingredient weight ratio (lb/lb) of about 1:1 (bacterial strain synthetic fungicide) to about 50: 1, about 1 : 1 to about 40: 1, about 10: 1 to about 40: 1, or about 10: 1 to about 30:1, including but not limited to, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26:1, about 27:1, about 28:1, about 29:1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35.1:1, about 36:1, about 37:1, about 38:1, about 39:1,
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with chlorothalonil in an active ingredient weight ratio (lb/lb) of about 1 : 100 to about 500: 1, including but not limited to, about 1 : 100, about 1:50, about 1:10, about 1:1, about 10:1, about 50: 1, about 100:1, about 150:1, about 200:1, about 250:1, about 300:1, about 350:1, about 400:1, about 450:1, and about 500:1.
  • active ingredient weight ratio lb/lb
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with chlorothalonil in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial strain: synthetic fungicide) to about 10:1, including but not limited to, about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1.9, about 1:1.8, about 1:1.75, about 1:1.7, about 1:1.65, about 1:1.6, about 1:1.55, about 1:1.5, about 1:1.45, about 1:4, about 1:1.35, about 1:1.3, about 1:1.275, about 1:1.25, about 1:1.24, about 1:1.23, about 1:1.22, about 1:1.21, about 1:1.2, about 1:1.19, about 1:1.18, about 1:1.17, about 1:1.16, about 1:1.15, about 1:1.15, about 1:1.1, about 1:1, about 1.1:1, about 1.2:1, about 1.3:1, about 1.4:
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with triflumizole in an active ingredient weight ratio (lb/lb) of about 1:10 (bacterial straimtriflumizole) to about 100: 1, including but not limited to, about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6: 1, about 7:1, about 8:1, about 9:1, about 10: 1, about 15:1, about 20: 1, about 25:1, about 30: 1, about 35:1, about 40:1, about 45:1, about 50:1, about 55:1, about 60:1, about 65:1, about 70:1, about 75:1, about 80: 1, about 85:1, about 90: 1, about 95:1, or about 100: 1
  • the bacterial strain, or active variant thereof can be applied (simultaneously or sequentially) with triflumizole in an active ingredient weight ratio (
  • Any appropriate agricultural application rate for a biocide can be applied (simultaneously or sequentially) with the combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and bacterial strain provided herein or active variant thereof disclosed herein.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and bacterial strain provided herein or active variant thereof disclosed herein.
  • Methods to assay for the effective amount of the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) and bacterial strain provided herein or active variant thereof include, for example, the control of the pathogen or pest targeted by the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) and bacterial strain combination. Methods to assay for such control are known.
  • a control of plant health, yield and/or growth that occurs upon application of an effective amount of the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) and a bacterial strain provided herein or active variant thereof.
  • the synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole
  • a method for controlling or inhibiting the growth of a plant pathogen that causes plant disease by applying (simultaneously or sequentially) at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain provided herein or active variant thereof provided herein (i.e., AIPI620, AIP050999, and CGA267356, or an active variant or any thereof, or a combination of cells from any one of AIP1620, AIP05G999, and CGA267356, or an active variant any of thereof).
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one
  • By“applying” is intended contacting an effective amount of the synthetic fungicide and a bacterial strain provided herein or active variant thereof to a plant, area of cultivation, seed and/or weed with one or more of the bacterial strains provided herein or active variant thereof so that a desired effect is achieved.
  • the application of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and/or the bacterial strain provided herein or active variant thereof can occur prior to the planting of the crop (for example, to the soil, the seed, or the plant).
  • the application of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and/or the bacterial strain provided herein or active variant thereof is a foliar application.
  • a further embodiment of the invention provides a method for controlling or inhibiting the growth of a plant pathogen by applying (simultaneously or sequentially) at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and the population of bacterial strain provided herein or active variant thereof to an environment in which the plant pathogen may grow.
  • the application may be to the plant, to parts of the plant, to the seeds of the plants to be protected, or to the soil in which the plant to be protected are growing or will grow.
  • Application to the plant or plant parts may be before or after harvest. Application to the seeds will be prior to planting of the seeds.
  • Application (simultaneous or sequential) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole can occur before, during, or after application of a bacterial strain provided herein, or active variant thereof.
  • application of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole is alternated with application of a bacterial strain provided herein, or active variant thereof.
  • Application of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole can alternate with application of a bacterial strain provided herein, or active variant thereof, by 0.5 days, 1 day, 1.5 days, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 14 days or more.
  • At least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole is applied to a plant, plant part, or cultivation area of a plant, simultaneously with a bacterial strain provided herein, or active variant thereof.
  • Simultaneous application of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and a bacterial strain provided herein, or active variant thereof, can occur in the same formulation or simultaneously from separate formulations.
  • At least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and a bacterial strain provided herein, or active variant thereof are formulated in the same composition to be applied as a foliar treatment in the same formulation at an effective amount less than the standard rate of application used for the synthetic fungicide (i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difenoconazole) or the bacterial strain AIP1620, or an active variant thereof, alone.
  • the synthetic fungicide i.e., tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, or difen
  • Sequential application of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and a bacterial strain provided herein, or active variant thereof refers to the application of each of the synthetic fungicide and bacterial strain that does not occur simultaneously.
  • Sequential application of two components includes the application of component a) before or after component b), wherein the difference in time between application of components a) and b) is greater than 1 minute, including but not limited to 1 minute, 5 minutes, 30 minutes,
  • timing of the sequential application of the synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and the bacterial strain provided herein, or active variant thereof is such that an additive effect or synergistic effect (on the controlling of a plant pathogen or treatment or prevention of a plant disease, or an improvement of an agronomic trait) is observed.
  • an effective amount of a combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain provided herein or active variant thereof provided herein is used as a foliar application to control or inhibit growth of one or more pathogens selected from the group consisting of Alternaria spp., Alternaria solani, Colletotrichum spp.
  • Mycosphaerella spp. Mycosphaerella spp., Phomopsis spp., Pseudoperonospora spp., Podosphaera spp., Cercospora spp., Corynespora spp., Gleocercospora spp., Sclerotinia spp., Bremia spp., Botrytis spp., and Botrytis cinerea.
  • an effective amount of a combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain provided herein or active variant thereof provided herein is applied (simultaneously or sequentially) to the soil in which the plant to be protected are growing or will grow to control or inhibit growth of one or more pathogens selected from the group consisting of Rhizoctonia spp., Rhizoctonia solani, Fusarium spp., Sclerotium spp., Sclerotinia spp., Sclerotinia sclerotiorum, Phytopthora spp., and Pythium spp.
  • an effective amount of a combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain provided herein or active variant thereof provided herein is applied (simultaneously or sequentially) to the plant after harvest to control or inhibit growth of one or more pathogens selected from the group consisting of Monolinia spp., Penici Ilium spp., Botrytis ssp., and Botrytis cinerea.
  • the term plant includes plant cells, plant protoplasts, plant cell tissue cultures from which plants can be regenerated, plant calli, plant clumps, and plant cells that are intact in plants or parts of plants such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruit, kernels, ears, cobs, husks, stalks, roots, root tips, anthers, and the like.
  • Grain is intended to mean the mature seed produced by commercial growers for purposes other than growing or reproducing the species.
  • a plant or plant part having an effective amount of a combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and the bacterial strain provided herein or active variant thereof (i.e., AIP1620, AIP050999, and CGA267356, or an active variant of any thereof, or a combination of cells), applied to the surface of the plant or plant part.
  • the combination of bacterial strain, or active variant thereof, and synthetic fungicide applied to the surface of the plant or plant part can be in the form a composition or formulation as disclosed elsewhere herein.
  • an effective amount of the combination is applied to a plant that has been removed from the field or area of cultivation or applied to a plant part that has been removed from a plant.
  • en effective amount of the combination disclosed herein can be applied to the surface of a seed of a plant.
  • the effective amount of the combination comprises a bacterial strain provided herein or active variant thereof in combination with a synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole in a weight ratio (lb/lb) disclosed herein for the selected synthetic fungicide.
  • the combination comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells (e.g., bacterial cells in the combination) on the surface of the plant or plant part.
  • chlorothalonil, and difenoconazole and the bacterial strain provided herein or active variant thereof i.e., AGR162.0, AIP050999, and CGA267356, or an active variant of any thereof, or a combination of cells, is applied (simultaneously or sequentially) to the leaves of a soybean plant.
  • the timing of application can vary depending on the conditions and geographical location.
  • the combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and the bacterial strain provided herein or active variant thereof is applied at the R1 (beginning flowering stage) of soybean development or may be applied earlier depending on ASR onset and the disease severity.
  • Such methods can comprise controlling a plant pathogen or population thereof in an area of cultivation by contacting the plant pathogen or population thereof with a combination (applied simultaneously or sequentially) of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain provided herein or active variant thereof (i.e., AIP1620, AIP050999, and CGA267356, or an active derivative or any thereof, or a combination of cells from any one of AIP1620, AIP050999, and CGA267356, or an active variant thereof).
  • a synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at
  • a method for controlling a plant pathogen in an area of cultivation can comprise planting the area of cultivation with seeds or plants susceptible to the plant disease; and applying (simultaneously or sequentially) to the plant susceptible to the disease, the seed or the area of cultivation of the plant susceptible to the plant disease an effective amount of a combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain provided herein or active variant thereof (i.e., AIP1620, AIP050999, and
  • CGA267356 or an active derivative or any thereof, or a combination of cells from any one of AIP1620, AIP050999, and CGA267356, or an active variant thereof), wherein the combination controls the plant disease without significantly affecting the crop or plant.
  • the method comprises applying (simultaneously or sequentially) to a plant susceptible to the disease, a seed, or an area of cultivation of the plant susceptible to the disease an effective amount of a combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain provided herein or active variant thereof.
  • at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain provided herein or active variant thereof.
  • the bacterial strain provided herein or active variant thereof may comprise at least one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof; or a combination of cells from any one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof.
  • an effective amount of the combination comprises a weight ratio (lb/lb) disclosed herein for the selected synthetic fungicide.
  • the application rate of the synthetic fungicide disclosed herein can be calculated based on the application rate of the bacterial strain, or active varian thereof, according to the selected active ingredient weight ratio as disclosed elsewhere herein for each selected synthetic fungicide.
  • an effective amount of the combination comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells (e.g., bacterial cells in the combination).
  • Methods are provided for controlling a plant pathogen or plant pest on a plant or plant part by applying to the plant or plant part an effective amount of a combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and the bacterial strain provided herein or active variant thereof (i.e., AGR162.0, A ⁇ R050999, and CGA267356, or an active variant of any thereof, or a combination of cells).
  • the composition comprising the bacterial strain or active variant thereof can be a solid or liquid composition or formulation.
  • the plant or plant part need not be actively growing in order for the bacterial strain to effectively control the plant pathogen or plant pest.
  • the plant or plant part has been harvested or otherwise removed from the field or area of cultivation.
  • An effective amount of the combination of bacterial strain, or active variant thereof, and synthetic fungicide can be applied to a plant or plant part prior to harvesting or after the plant or plant part has been harvested.
  • harvesting refers to the removal of a plant or plant part from the ground or other area of cultivation and can also refer to removal of a plant part from a plant that remains in the ground or other area of cultivation.
  • the effective amount of the combination comprises a bacterial strain provided herein or active variant thereof in combination with a synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole in a weight ratio (lb/lb) disclosed herein for the selected synthetic fungicide.
  • an effective amount of the combination comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells (e.g., bacterial cells in the combination).
  • the “yield” of the plant refers to the quality and/or quantity of biomass produced by the plant.
  • biomass is intended any measured plant product.
  • An increase in biomass production is any improvement in the yield of the measured plant product.
  • An increase in yield can comprise any statistically significant increase including, but not limited to, at least a 1% increase, at least a 3% increase, at least a 5% increase, at least a 10% increase, at least a 20% increase, at least a 30%, at least a 50%, at least a 70%, at least a 100% or a greater increase in yield compared to a plant not exposed to the bacterial strain provided herein or active variant thereof.
  • a method for increasing yield in a plant comprises applying (simultaneously or sequentially) to a crop, a plant, or an area of cultivation an effective amount of a combination of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and at least one bacterial strain comprising AIP1620, AIP050999, and CGA267356, or an active variant of any thereof, or a combination of cells from any one of AIP1620, AIP050999, and
  • an effective amount of the combination comprises at least about 400 pg pyrrolnitrin per gram of bacterial cells (e.g., bacterial cells in the combination).
  • an“area of cultivation” comprises any region in which one desires to grow a plant.
  • Such areas of cultivations include, but are not limited to, a field in which a plant is cultivated (such as a crop field, a plant field, a sod field, a tree field, a managed forest, a field for culturing fruits and vegetables, etc.), a greenhouse, a growth chamber, etc.
  • a plant of interest i.e., plant susceptible to the plant disease
  • the area of cultivation comprising the plant can be treated with an effective amount of a combination (applied simultaneously or sequentially) of a bacterial strain provided herein or active variant thereof and at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole.
  • a bacterial strain provided herein or active variant thereof and a biocide By“treated with a combination of’ or“applying a combination of’ a bacterial strain provided herein or active variant thereof and a biocide to a plant, area of cultivation or field it is intended that one or more of a particular field, plant, and/or weed is treated (applied simultaneously or sequentially) with an effective amount of one or more of the bacterial strain provided herein or active variant thereof and one or more biocide so that a desired effect is achieved.
  • the application of one or both of the bacterial strain provided herein or active variant thereof and the biocide can occur prior to the planting of the crop (for example, to the soil, or the plant).
  • the application of the bacterial strain provided herein or active variant thereof and the biocide may be simultaneous or the applications may be at different times (sequential), so long as the desired effect is achieved.
  • the active variant comprises a bacterial strain provided herein that is resistant to one or more biocide.
  • the bacterial strain provided herein or active variant thereof i.e., AIP1620, AIP050999, and CGA267356, or an active variant of any thereof, or a combination of cells from any one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof
  • a plant, crop, or area of cultivation is treated
  • the active variant comprises a bacterial strain provided herein that is resistant to glufosinate.
  • a plant, crop, or area of cultivation is treated
  • the effective amount of at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole and the bacterial strain provided herein or active variant thereof is sufficient to result in an improvement in plant health, yield, and/or growth when compared to the plant health, yield, and/or growth that occurs when the same concentration of the synthetic fungicide and bacterial strain provided herein or active variant thereof that was not modified to be resistant to glufosinate is applied in combination with the effective amount of the glufosinate or active derivative thereof.
  • the effective amount comprises a bacterial strain provided herein or active variant thereof such as AIP1620, AIP050999, and CGA267356, or an active variant of any thereof, or a combination of cells from any one of AIP1620, AIP050999, and CGA267356, or an active variant of any thereof.
  • the bacterial strain provided herein or active variant thereof can be used in combination (applied simultaneously or sequentially) with at least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole,
  • chlorothalonil, and difenoconazole and another biocide i.e., a herbicide, fungicide, pesticide, or other crop protection chemical.
  • another biocide i.e., a herbicide, fungicide, pesticide, or other crop protection chemical.
  • the bacterial strain provided herein or active variant thereof is compatible with the additional biocide of interest.
  • herbicide, fungicide, pesticide, or other crop protection chemical tolerance or herbicide, fungicide, pesticide, or other crop protection chemical resistance is intended the ability of an organism (i.e., the plant and/or the bacterial strain provided herein or active variant thereof etc.) to survive and reproduce following exposure to a dose of the herbicide, fungicide, pesticide, or other crop protection chemical that is normally lethal to the wild type organism.
  • Herbicides that can be used in the various methods and compositions disclosed herein include glyphosate, ACCase inhibitors (Arloxyphenoxy propionate (FOPS)); ALS inhibitors (Sulfonylurea (SU)), Imidazonlinone (IMI), Pyrimidines (PM)); microtubule protein inhibitor (Dinitroaniline (DNA)); synthetic auxins (Phenoxy (P)), Benzoic Acid (BA), Carboxylic acid (CA)); Photosystem II inhibitor (Triazine (TZ)), Triazinone (TN), Nitriles (NT), Benzothiadiazinones (BZ), Ureas (US)); EPSP Synthase inhibitor (glycines (GC)); Glutamine Synthesis inhibitor (Phosphinic Acid (PA)); DOXP synthase inhibitor (Isoxazolidinone (IA)); HPPD inhibitor (Pyrazole (PA)), Triketone (TE)); PPO inhibitors (
  • Pesticides that can be used in the various methods and compositions disclosed herein include imidacloprid clothianidin, arylpyrazole compounds (W02007103076); organophosphates, phenyl pyrazole, pyrethoids caramoyloximes, pyrazoles, amidines, halogenated hydrocarbons, carbamates and derivatives thereof, terbufos, chloropyrifos, fipronil, chlorethoxyfos, telfuthrin, carbofuran, imidacloprid, tebupirimfos (U.S. Patent No. 5,849,320).
  • Additional fungicides that can be used in the various methods and compositions disclosed herein include aliphatic nitrogen fungicides (butylamine, cymoxanil, dodicin, dodine, guazatine, iminoctadine); amide fungicides (benzovindiflupyr, carpropamid, chloraniformethan, cyflufenamid, diclocymet, diclocymet, dimoxystrobin, fenaminstrobin, fenoxanil, flumetover, furametpyr, isofetamid, isopyrazam, mandestrobin, mandipropamid, metominostrobin, orysastrobin, penthiopyrad, prochloraz, quinazamid, silthiofam, triforine); acylamino acid fungicides (benalaxyl, benalaxyl-M, furalaxyl, metalaxyl, metalaxyl- M,
  • sulfonamide fungicides (amisulbrom, cyazofamid); valinamide fungicides (benthiavalicarb, iprovalicarb); antibiotic fungicides (aureofungin, blasticidin-S, cycloheximide, griseofulvin, kasugamycin, moroxydine, natamycin, polyoxins, polyoxorim, streptomycin, validamycin); strobilurin fungicides (fluoxastrobin, mandestrobin); other methoxyacrylate strobilurin fungicides (bifujunzhi, coumoxystrobin, enoxastrobin, flufenoxystrobin, jiaxiangjunzhi, picoxystrobin, pyraoxystrobin); methoxycarbanilate strobilurin fungicides (pyraclostrobin, pyrametostrobin, triclopyricarb); methoxyiminoace
  • strobin fenaminstrobin, metominostrobin, orysastrobin
  • methoxyiminoacetate strobilurin fungicides kresoxim-methyl, trifloxystrobin
  • other aromatic fungicides biphenyl
  • chlorodinitronaphthalenes chloroneb, cresol, dicloran, fenjuntong, hexachlorobenzene, pentachlorophenol, quintozene, sodium pentachlorophenoxide, tecnazene, trichlorotrinitrobenzenes); arsenical fungicides (asomate, urbacide); aryl phenyl ketone fungicides (metrafenone, pyriofenone); benzimidazole fungicides (albendazole, benomyl, carbendazim, chlorfenazole, cypendazole, debacarb, fuberidazole, mecarbinzid, rabenzazole, thiabendazole); benzimidazole precursor fungicides (furophanate, thiophanate, thiophanate- methyl); benzothiazole fungicides (bentaluron, benthiavalicarb
  • hydrazide fungicides (benquinox, saijunmao); imidazole fungicides (cyazofamid, fenamidone, fenapanil, glyodin, iprodione, isovaledione, pefurazoate, triazoxide); conazole fungicides (imidazoles) (climbazole, clotrimazole, imazalil, oxpoconazole, prochloraz, triflumizole); inorganic fungicides (potassium azide, potassium thiocyanate, sodium azide, sulfur, see also copper fungicides, see also inorganic mercury fungicides); mercury fungicides; inorganic mercury fungicides (mercuric chloride, mercuric oxide, mercurous chloride); organomercury fungicides ((3-ethoxypropyl)mercury bromide, ethy
  • organotin fungicides decafentin, fentin, tributyltin oxide
  • oxathiin fungicides carboxin, oxycarboxin
  • oxazole fungicides chlorozolinate, dichlozoline, drazoxolon, famoxadone, hymexazol, metazoxolon, myclozolin, oxadixyl, oxathiapiprolin, pyrisoxazole, vinclozolin
  • polysulfide fungicides barium polysulfide
  • fungicides acibenzolar, acypetacs, allyl alcohol, benzalkonium chloride, bethoxazin, bromothalonil, chitosan, chloropicrin, DBCP, dehydroacetic acid, diclomezine, diethyl pyrocarbonate, ethylicin, fenaminosulf, fenitropan, fenpropidin, formaldehyde, furfural, hexachlorobutadiene, methyl isothiocyanate, nitrostyrene, nitrothal-isopropyl, OCH, pentachlorophenyl laurate, 2-phenylphenol, phthalide, piperalin, propamidine, proquinazid, pyroquilon, sodium
  • Non-limiting embodiments of the invention include:
  • composition comprising:
  • At least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole;s
  • composition comprises an active ingredient weight ratio selected from the group consisting of:
  • composition of embodiment 1, wherein said composition comprises an active ingredient weight ratio selected from the group consisting of:
  • composition of embodiment 2, wherein said composition comprises an active ingredient weight ratio selected from the group consisting of:
  • composition of any one of embodiments 1-4 wherein said composition comprises a wettable powder, dry formulation, wettable granule, spray dried formulation, or agglomerated formulation comprising said bacterial strain or the active variant of any thereof.
  • composition of embodiment 5, wherein said composition comprises a dry flowable comprising at least one of azoxystrobin, chlorothalonil, flutriafol, triflumazole, difenconazole, and tebuconazole.
  • composition of embodiment 5, wherein said composition comprises a liquid flowable formulation comprising at least one of azoxystrobin, chlorothalonil, flutriafol, triflumazole, and
  • composition of any one of embodiments 1-9 wherein an effective amount of the composition improves an agronomic trait of interest of a plant or controls a plant pathogen that causes a plant disease.
  • composition of embodiment 10, wherein the plant disease is a plant disease caused by a fungal pathogen or fungal-like pathogen.
  • composition of embodiment 10 wherein the plant pathogen is Asian Soybean Rust (ASR), powdery mildew, anthracnose, target spot, early blight, head drop, downy mildew, gray mold, and/or zonate leaf spot.
  • ASR Asian Soybean Rust
  • said plant pathogen comprises at least one fungal pathogen or fungal-like pathogen.
  • composition of embodiment 10, wherein said at least one plant pathogen comprises Botrytis spp.. Bremia spp., Cersospora spp., Corynespora spp., Alternatia spp., Fusarium spp., Podosphaera spp., Gleocercospora spp., Pseudoperonospora spp., Phakopsora sp., Puccini a spp., Pythium spp., Phytophthora spp., Rhizoctonia spp., Sclerotinia spp., Verticillium spp., Colletotrichum ssp. or Monilinia spp..
  • composition of embodiment 13, wherein said at least one plant pathogen comprises Botrytis cinerea, Cercospora sojina, Corynespora cassiicola, Alternaria solani, Alternaria dauci, Bremia lactucae, Gleocercospora sorghi, Rhizoctonia solani, Erysiphe necator, Podosphaera xanthii,
  • Colletotrichum cereal Colletotrichum graminicola, Fusarium nivale, Plasmopara viticola, Peronospora helhahrii, Pythium aphanidermatum, Pythium sylvaticum, Pythium myriotylum, Pythium ultimum,
  • composition of any one of embodiments 1-16 wherein said composition comprises at least one of AIP1620, AIP050999, and CGA267356, or the active variant of any thereof at a concentration for application lower than the suggested application rate.
  • composition of embodiment 17, wherein said composition comprises at least one of AIP1620, AIP050999, and CGA267356, or the active variant of any thereof at a concentration for application at about 0.5 lb/acre to about 2.49 lb/acre.
  • composition of embodiment 18, wherein said composition comprises at least one of AIP1620, AIP050999, and CGA267356, or the active variant of any thereof at a concentration for application at about 0.5 lb/acre or about 1.25 lb/acre.
  • compositions of any one of embodiments 1-19 wherein said composition comprises a kit having said synthetic fungicide in a spatially separated arrangement from said at least one of AIP1620, AIP050999, and CGA267356, or the active variant of any thereof.
  • a plant or plant part comprising an effective amount of the composition of any one of embodiments 1-20 on the surface of said plant or plant part.
  • At least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole;
  • said method comprises applying (a) and (b) at an active ingredient weight ratio selected from the group consisting of:
  • said effective amount controls a plant pathogen that causes the plant disease or improves the agronomic trait of interest.
  • a method for controlling a plant pathogen that causes a plant disease in an area of cultivation comprising:
  • a combination comprising: (a) at least one of bacterial strainAIP1620, A IPO 50999. and CGA267356, or an active variant of any thereof; and wherein the active variant comprises a bacterial strain having a genome within a Mash distance of about 0.015; and
  • At least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole;
  • said method comprises applying (i) and (ii) at an active ingredient weight ratio selected from the group consisting of:
  • said effective amount controls said plant pathogen that causes said plant disease.
  • At least one synthetic fungicide selected from the group consisting of tetraconazole, triflumizole, azoxystrobin, flutriafol, tebuconazole, chlorothalonil, and difenoconazole;
  • said method comprises applying (a) and (b) at an active ingredient weight ratio selected from the group consisting of:
  • said effective amount controls a plant pathogen that causes the plant disease.
  • CFU colony forming units
  • said at least one plant pathogen comprises Botrytis spp., Bremia spp., Cersospora spp., Corynespora spp., Alternatia spp., Fusarium spp., Podosphaera spp., Gleocercospora spp., Pseudoperonospora spp., Phakopsora sp., Puccini a spp., Pythium spp.,
  • Phytophthora spp. Rhizoctonia spp., Sclerotinia spp., Verticillium spp., Colletotrichum ssp., andMonilinia spp..
  • said plant pathogen comprises Botrytis cinerea, Cercospora sojina, Corynespora cassiicola, Alternaria solani, Alternaria dauci, Bremia lactucae,
  • Gleocercospora sorghi Rhizoctonia solani, Erysiphe necator, Podosphaera xanthii, Colletotrichum cereal, Colletotrichum graminicola, Fusarium nivale, Plasmopara viticola, Peronospora helhahrii, Pythium aphanidermatum, Pythium sylvaticum, Pythium myriotylum, Pythium ultimum, Phytophthora nicotianae, Phytophthora infestans, Phytophthora tropicalis, Phytophthora sojae, Pseudoperonospora cubensis, Fusarium graminearum, Fusarium solani, Phakopsora pachyrizi, Sclerotinia minor, or Venturia inaequalisa.
  • the target crop was grown in research plots using normal production methods. Products were applied as a foliar spray in water along with a non-ionic surfactant several times during the growing season. AIP1620 was applied at a half rate of 2.5 lbs/acre alone, or in combination with a commercially-available formulation of each synthetic fungicide. Products were mixed in spray tanks and applied to four replicate plots as a foliar spray. Unless indicated otherwise, natural infestation of the crop occurred. Disease incidence or severity was visually evaluated periodically, depending on the level of infestation and particular disease. Results across replicated plots were averaged to account for natural variability. Treated plots were compared to untreated controls and Percent Control for each treatment was calculated as follows:
  • AIP1620 and azoxystrobin were tested on target spot of tomato ( Corynespora cassiicola). Disease severity was measured and results are provided in Table 2. Synergy between AIP1620 and azoxystrobin was observed.
  • AIP1620 and azoxystrobin were tested on early blight of tomato ( Alternaria solani). Disease severity was measured and results are provided in Table 3. Synergy between AIP1620 and azoxystrobin was observed.

Abstract

L'invention concerne des compositions et des procédés pour le traitement ou la prévention de maladies des plantes. Ces compositions et procédés comprennent une combinaison d'un fongicide synthétique sélectionné dans le groupe constitué par le tétraconazole, le triflumizole, l'azoxystrobine, le flutriafol, le tébuconazole, le chlorothalonil et le difénoconazole et une souche bactérienne qui lutte contre au moins un pathogène qui provoque une maladie des plantes ou améliore au moins un caractère agronomique d'intérêt dans une plante. La combinaison du fongicide synthétique et de la souche bactérienne peut être utilisée comme inoculant pour des plantes. Par conséquent, l'invention concerne également des procédés de culture d'une plante sensible à une maladie des plantes et des procédés de lutte contre une maladie des plantes sur une plante sensible à la maladie des plantes.
EP20750436.6A 2019-06-07 2020-06-05 Compositions et procédés d'amélioration de la santé des plantes et de lutte contre les maladies des plantes Pending EP3979802A1 (fr)

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CN114173562A (zh) 2022-03-11
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BR112021024728A2 (pt) 2022-05-03
ECSP22001411A (es) 2022-04-29
US20220312774A1 (en) 2022-10-06
PE20220899A1 (es) 2022-05-30
MX2021015088A (es) 2022-01-18
WO2020247848A1 (fr) 2020-12-10
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KR20220031608A (ko) 2022-03-11

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